{"database": "metadata", "table": "run_metadata", "is_view": false, "human_description_en": "where experiment.library_strategy = \"RNA-Seq\", technology = \"unknown\" and tissue_curation = \"Skin\"", "rows": [[3759, "ERR1294279", "ERX1365625", "ERS1067820", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVBU1/tVBU1 skin transcriptome", "SAMEA3880686", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880686|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VBU1|allele:mau tVBU1/tVBU1|common name:zebrafish|dev stage:adult|sample name:VBU1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 9", "vbu13", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "vbu13_GCCAAT_L008_R1_001.fastq.gz", "fastq", 2376143675.0, 23526175.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 9", "0:101", "A:655940769;C:534990399;G:527934646;T:655773043;N:1504818", 101, null, null, null, 655940769, 534990399, 527934646, 655773043, 1504818, "ERX1365625", "ERS1067820", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.95443, null, 0.10666, null, 0.73176, null, 0.50978, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3760, "ERR1294278", "ERX1365624", "ERS1067820", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVBU1/tVBU1 skin transcriptome", "SAMEA3880686", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880686|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VBU1|allele:mau tVBU1/tVBU1|common name:zebrafish|dev stage:adult|sample name:VBU1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 8", "vbu12", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "vbu12_GTGAAA_L008_R1_001.fastq.gz", "fastq", 1449181835.0, 14348335.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 8", "0:101", "A:399518584;C:327615685;G:323868050;T:397240465;N:939051", 101, null, null, null, 399518584, 327615685, 323868050, 397240465, 939051, "ERX1365624", "ERS1067820", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.95893, null, 0.09315, null, 0.73464, null, 0.48744, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3761, "ERR1294277", "ERX1365623", "ERS1067820", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVBU1/tVBU1 skin transcriptome", "SAMEA3880686", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880686|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VBU1|allele:mau tVBU1/tVBU1|common name:zebrafish|dev stage:adult|sample name:VBU1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 7", "vbu11", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "vbu11_GTCCGC_L008_R1_001.fastq.gz", "fastq", 1455235169.0, 14408269.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 7", "0:101", "A:392100064;C:337980617;G:331091626;T:393178582;N:884280", 101, null, null, null, 392100064, 337980617, 331091626, 393178582, 884280, "ERX1365623", "ERS1067820", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.95485, null, 0.08016, null, 0.72845, null, 0.47549, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3762, "ERR1294276", "ERX1365622", "ERS1067819", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVE1/tVE1 skin transcriptome", "SAMEA3880685", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880685|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VE1|allele:mau tVE1/tVE1|common name:zebrafish|dev stage:adult|sample name:VE1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 6", "ve13", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "ve13_CCGTCC_L008_R1_001.fastq.gz", "fastq", 1029294030.0, 10191030.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:670 6", "0:101", "A:284612731;C:231451012;G:228443320;T:284135536;N:651431", 101, null, null, null, 284612731, 231451012, 228443320, 284135536, 651431, "ERX1365622", "ERS1067819", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.93287, null, 0.10726, null, 0.72585, null, 0.4913, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3763, "ERR1294275", "ERX1365621", "ERS1067819", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVE1/tVE1 skin transcriptome", "SAMEA3880685", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880685|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VE1|allele:mau tVE1/tVE1|common name:zebrafish|dev stage:adult|sample name:VE1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 5", "ve12", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "ve12_ATGTCA_L008_R1_001.fastq.gz", "fastq", 1446322222.0, 14320022.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 5", "0:101", "A:400821160;C:324210104;G:320286425;T:400078485;N:926048", 101, null, null, null, 400821160, 324210104, 320286425, 400078485, 926048, "ERX1365621", "ERS1067819", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.92778, null, 0.09489, null, 0.73093, null, 0.48493, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3764, "ERR1294274", "ERX1365620", "ERS1067819", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "Mau tVE1/tVE1 skin transcriptome", "SAMEA3880685", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880685|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:VE1|allele:mau tVE1/tVE1|common name:zebrafish|dev stage:adult|sample name:VE1|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 4", "ve11", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "ve11_AGTTCC_L008_R1_001.fastq.gz", "fastq", 2117238861.0, 20962761.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 4", "0:101", "A:591654664;C:469803769;G:462550117;T:591863795;N:1366516", 101, null, null, null, 591654664, 469803769, 462550117, 591863795, 1366516, "ERX1365620", "ERS1067819", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.93146, null, 0.09666, null, 0.73501, null, 0.48749, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3765, "ERR1294273", "ERX1365619", "ERS1067818", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "WT skin transcriptome", "SAMEA3880684", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880684|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:WT|allele:mau +/+|common name:zebrafish|dev stage:adult|sample name:WT|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 3", "wt3", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "wt3_AGTCAA_L008_R1_001.fastq.gz", "fastq", 2631687411.0, 26056311.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 3", "0:101", "A:734843629;C:584316000;G:577487406;T:733356850;N:1683526", 101, null, null, null, 734843629, 584316000, 577487406, 733356850, 1683526, "ERX1365619", "ERS1067818", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.95114, null, 0.1036, null, 0.72474, null, 0.48261, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3766, "ERR1294272", "ERX1365618", "ERS1067818", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "WT skin transcriptome", "SAMEA3880684", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880684|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:WT|allele:mau +/+|common name:zebrafish|dev stage:adult|sample name:WT|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 2", "wt2", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "wt2_CTTGTA_L008_R1_001.fastq.gz", "fastq", 2093443463.0, 20727163.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:669 2", "0:101", "A:607582034;C:442802229;G:437957089;T:603752294;N:1349817", 101, null, null, null, 607582034, 442802229, 437957089, 603752294, 1349817, "ERX1365618", "ERS1067818", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.94792, null, 0.10892, null, 0.73277, null, 0.51067, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [3767, "ERR1294271", "ERX1365617", "ERS1067818", "ERP014370", "PRJEB12848", "Danio rerio Mau mutants skin Transcriptome", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-25-02-2016-18:42:00:594-265", "Other", "Mau/Aqp3a dominant mutants of zebrafish Danio rerio are characterized by broken stripes and short fins. To identify signaling pathways affected by mau mutations and leading to pigment patterning defects  we analyze skin transcriptome of adult wild type TU fish and two mau alleles.", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2016 02 25", null, null, "WT skin transcriptome", "SAMEA3880684", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2016 05 24|ENA last update:2016 02 25|External Id:SAMEA3880684|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2016 05 24T17:02:01Z|INSDC last update:2016 02 25T18:42:06Z|INSDC status:public|Submitter Id:WT|allele:mau +/+|common name:zebrafish|dev stage:adult|sample name:WT|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina HiSeq 2000 sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:668 1", "wt1", "1", "Illumina TruSeq RNA", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "ERP014370", "Illumina HiSeq 2000 sequencing", "ENA FIRST PUBLIC:2016 05 24|ENA LAST UPDATE:2018 11 16", "wt1_CAGATC_L008_R1_001.fastq.gz", "fastq", 1701561241.0, 16847141.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 25 02 2016 18:41:58:668 1", "0:101", "A:505358711;C:348893979;G:343445186;T:502880170;N:983195", 101, null, null, null, 505358711, 348893979, 343445186, 502880170, 983195, "ERX1365617", "ERS1067818", "ERA567249", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 1, 0.94565, null, 0.13032, null, 0.73669, null, 0.50473, null, 101, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2016-02-25", "Adult", "Adult", "Skin", "Surface Structure"], [9770, "ERR5838122", "ERX5487778", "ERS6337138", "ERP119543", "PRJEB36360", "Transcriptomic analysis of adult skin from 9 Danio species", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-22-01-2020-08:38:04:562-27", "Other", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "ENA FIRST PUBLIC:2020 03 21|ENA LAST UPDATE:2021 04 21", "PUBMED:33277491", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "Transcriptomic analysis of adult skin from 9 Danio species", "SAMEA8652584", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2021 04 30|ENA last update:2021 04 29|External Id:SAMEA8652584|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2021 04 30T00:30:20Z|INSDC last update:2021 04 29T02:45:00Z|INSDC status:public|Submitter Id:05 Danio rerio adult skin 5|collected by:Marco Podobnik|collection date:2019 08 23|common name:zebrafish|dev stage:adult|identified by:Marco Podobnik|isolation source:Tuebingen|sample name:05 Danio rerio adult skin 5|sex:male|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina NovaSeq 6000 paired end sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 5", "unspecified", "1", "TruSeq stranded mRNA Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "PAIRED", "ILLUMINA", "Illumina NovaSeq 6000", null, "ERP119543", "Illumina NovaSeq 6000 paired end sequencing", "ENA FIRST PUBLIC:2021 04 30|ENA LAST UPDATE:2021 04 29", "S1385Nr5.1.fastq.gz S1385Nr5.2.fastq.gz", "fastq fastq", 6912206316.0, 34306523.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 5", "0:100.74 1:100.74", "A:1853779551;C:1607929988;G:1658402611;T:1792045990;N:48176", 100, 100, null, null, 1853779551, 1607929988, 1658402611, 1792045990, 48176, "ERX5487778", "ERS6337138", "ERA4129786", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 2, 0.96095, 0.962, 0.08049, 0.07821, 0.70654, 0.70926, 0.51398, 0.51064, 101, 101, "B", "B", "biological fallback assumption", "illumina", "novaseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2020-03-21", "Adult", "Adult", "Skin", "Surface Structure"], [9771, "ERR5838121", "ERX5487777", "ERS6337136", "ERP119543", "PRJEB36360", "Transcriptomic analysis of adult skin from 9 Danio species", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-22-01-2020-08:38:04:562-27", "Other", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "ENA FIRST PUBLIC:2020 03 21|ENA LAST UPDATE:2021 04 21", "PUBMED:33277491", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "Transcriptomic analysis of adult skin from 9 Danio species", "SAMEA8652582", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2021 04 30|ENA last update:2021 04 29|External Id:SAMEA8652582|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2021 04 30T00:30:20Z|INSDC last update:2021 04 29T02:45:00Z|INSDC status:public|Submitter Id:04 Danio rerio adult skin 4|collected by:Marco Podobnik|collection date:2019 08 23|common name:zebrafish|dev stage:adult|identified by:Marco Podobnik|isolation source:Tuebingen|sample name:04 Danio rerio adult skin 4|sex:male|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina NovaSeq 6000 paired end sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 4", "unspecified", "1", "TruSeq stranded mRNA Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "PAIRED", "ILLUMINA", "Illumina NovaSeq 6000", null, "ERP119543", "Illumina NovaSeq 6000 paired end sequencing", "ENA FIRST PUBLIC:2021 04 30|ENA LAST UPDATE:2021 04 29", "S1385Nr4.1.fastq.gz S1385Nr4.2.fastq.gz", "fastq fastq", 6161942050.0, 30595009.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 4", "0:100.70 1:100.70", "A:1642716052;C:1446485458;G:1490121106;T:1582576605;N:42829", 100, 100, null, null, 1642716052, 1446485458, 1490121106, 1582576605, 42829, "ERX5487777", "ERS6337136", "ERA4129786", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 2, 0.96544, 0.96564, 0.07368, 0.07303, 0.70822, 0.71092, 0.50154, 0.50781, 101, 101, "B", "B", "biological fallback assumption", "illumina", "novaseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2020-03-21", "Adult", "Adult", "Skin", "Surface Structure"], [9772, "ERR5838120", "ERX5487776", "ERS6337133", "ERP119543", "PRJEB36360", "Transcriptomic analysis of adult skin from 9 Danio species", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-22-01-2020-08:38:04:562-27", "Other", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "ENA FIRST PUBLIC:2020 03 21|ENA LAST UPDATE:2021 04 21", "PUBMED:33277491", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "Transcriptomic analysis of adult skin from 9 Danio species", "SAMEA8652579", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2021 04 30|ENA last update:2021 04 29|External Id:SAMEA8652579|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2021 04 30T00:30:20Z|INSDC last update:2021 04 29T02:45:00Z|INSDC status:public|Submitter Id:03 Danio rerio adult skin 3|collected by:Marco Podobnik|collection date:2019 08 23|common name:zebrafish|dev stage:adult|identified by:Marco Podobnik|isolation source:Tuebingen|sample name:03 Danio rerio adult skin 3|sex:female|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina NovaSeq 6000 paired end sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 3", "unspecified", "1", "TruSeq stranded mRNA Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "PAIRED", "ILLUMINA", "Illumina NovaSeq 6000", null, "ERP119543", "Illumina NovaSeq 6000 paired end sequencing", "ENA FIRST PUBLIC:2021 04 30|ENA LAST UPDATE:2021 04 29", "S1385Nr3.1.fastq.gz S1385Nr3.2.fastq.gz", "fastq fastq", 7422881394.0, 36866805.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 3", "0:100.67 1:100.67", "A:1950275527;C:1768648993;G:1827362519;T:1876542858;N:51497", 100, 100, null, null, 1950275527, 1768648993, 1827362519, 1876542858, 51497, "ERX5487776", "ERS6337133", "ERA4129786", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 2, 0.97031, 0.97047, 0.04915, 0.04817, 0.71459, 0.71894, 0.51089, 0.5097, 101, 101, "B", "B", "biological fallback assumption", "illumina", "novaseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2020-03-21", "Adult", "Adult", "Skin", "Surface Structure"], [9773, "ERR5838119", "ERX5487775", "ERS6337132", "ERP119543", "PRJEB36360", "Transcriptomic analysis of adult skin from 9 Danio species", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-22-01-2020-08:38:04:562-27", "Other", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "ENA FIRST PUBLIC:2020 03 21|ENA LAST UPDATE:2021 04 21", "PUBMED:33277491", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "Transcriptomic analysis of adult skin from 9 Danio species", "SAMEA8652578", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2021 04 30|ENA last update:2021 04 29|External Id:SAMEA8652578|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2021 04 30T00:30:20Z|INSDC last update:2021 04 29T02:45:00Z|INSDC status:public|Submitter Id:02 Danio rerio adult skin 2|collected by:Marco Podobnik|collection date:2019 08 23|common name:zebrafish|dev stage:adult|identified by:Marco Podobnik|isolation source:Tuebingen|sample name:02 Danio rerio adult skin 2|sex:female|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina NovaSeq 6000 paired end sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 2", "unspecified", "1", "TruSeq stranded mRNA Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "PAIRED", "ILLUMINA", "Illumina NovaSeq 6000", null, "ERP119543", "Illumina NovaSeq 6000 paired end sequencing", "ENA FIRST PUBLIC:2021 04 30|ENA LAST UPDATE:2021 04 29", "S1385Nr2.1.fastq.gz S1385Nr2.2.fastq.gz", "fastq fastq", 7006718028.0, 34786110.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 2", "0:100.71 1:100.71", "A:1847371523;C:1665346769;G:1712373266;T:1781565768;N:60702", 100, 100, null, null, 1847371523, 1665346769, 1712373266, 1781565768, 60702, "ERX5487775", "ERS6337132", "ERA4129786", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 2, 0.96136, 0.96039, 0.05872, 0.05752, 0.70325, 0.70674, 0.46717, 0.48218, 101, 101, "B", "B", "biological fallback assumption", "illumina", "novaseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2020-03-21", "Adult", "Adult", "Skin", "Surface Structure"], [9774, "ERR5838118", "ERX5487774", "ERS6337063", "ERP119543", "PRJEB36360", "Transcriptomic analysis of adult skin from 9 Danio species", "ena-STUDY-MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY-22-01-2020-08:38:04:562-27", "Other", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "ENA FIRST PUBLIC:2020 03 21|ENA LAST UPDATE:2021 04 21", "PUBMED:33277491", "Transcriptomic analysis was carried out on adult skin of the flanks for the following nine Danio species: Danio rerio  Danio aesculapii  Danio nigrofasciatus  Danio tinwini  Danio kyathit  Danio albolineatus  Danio choprae  Danio margaritatus and Danio erythromicron. Five replicates per species were carried out. Each replicate was derived from a single fish with skin samples coming from both flanks. Paired end  stranded RNA Seq was then carried out.", "Transcriptomic analysis of adult skin from 9 Danio species", "SAMEA8652509", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", "ENA first public:2021 04 30|ENA last update:2021 04 29|External Id:SAMEA8652509|INSDC center alias:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC center name:MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|INSDC first public:2021 04 30T00:30:20Z|INSDC last update:2021 04 29T02:45:00Z|INSDC status:public|Submitter Id:01 Danio rerio adult skin 1|collected by:Marco Podobnik|collection date:2019 08 23|common name:zebrafish|dev stage:adult|identified by:Marco Podobnik|isolation source:Tuebingen|sample name:01 Danio rerio adult skin 1|sex:male|tissue type:skin", null, null, null, null, null, null, null, null, "Illumina NovaSeq 6000 paired end sequencing", "ena EXPERIMENT MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 1", "unspecified", "1", "TruSeq stranded mRNA Illumina", null, "RNA-Seq", "TRANSCRIPTOMIC", "Oligo-dT", "PAIRED", "ILLUMINA", "Illumina NovaSeq 6000", null, "ERP119543", "Illumina NovaSeq 6000 paired end sequencing", "ENA FIRST PUBLIC:2021 04 30|ENA LAST UPDATE:2021 04 29", "S1385Nr1.1.fastq.gz S1385Nr1.2.fastq.gz", "fastq fastq", 6586457266.0, 32719900.0, "ena RUN MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY 28 04 2021 08:27:34:651 1", "0:100.65 1:100.65", "A:1709547533;C:1593125315;G:1617088678;T:1666650824;N:44916", 100, 100, null, null, 1709547533, 1593125315, 1617088678, 1666650824, 44916, "ERX5487774", "ERS6337063", "ERA4129786", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY|European Nucleotide Archive", "MAX PLANCK INSTITUTE FOR DEVELOPMENTAL BIOLOGY", 2, 0.9531, 0.95366, 0.07763, 0.07567, 0.71384, 0.71634, 0.52409, 0.52475, 101, 101, "B", "B", "biological fallback assumption", "illumina", "novaseq_era", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2020-03-21", "Adult", "Adult", "Skin", "Surface Structure"], [25297, "SRR25764099", "SRX21486772", "SRS18719072", "SRP457109", "PRJNA1009810", "Live tracking of basal stem cells of the epidermis during growth  homeostasis  and injury response in zebrafish", "GSE241757", "Transcriptome Analysis", "Basal stem cells of the epidermis continuously differentiate into keratinocytes and replenish themselves via self renewal to maintain skin homeostasis. Numerous studies have attempted to reveal how basal cells undergo differentiation or self renewal; however  this has been hampered by a lack of robust basal cell markers and analytical platforms that allow single cell tracking. Here  we report integrin b4 itgb4 is a useful marker for basal cell labelling  irrespective of the body region  stage  and regenerative status. We employed Cre loxP recombination in combination with live cell tracking of single basal clones in the caudal fin and investigated the embryonic origin and behaviour of basal cells during fish growth and homeostasis. Although most basal cells  including those in fins  became quiescent in the adult stage  genetic cell ablation showed that basal cells were reactivated to either self renew or differentiate  depending on the injured cell type. Our findings provide a platform for quantitative in vivo imaging of basal stem cells applicable at wider stages and under various conditions and reveal unanticipated basal cell dynamism in response to distinct wound signals. Overall design: To investigate whether itgb4+ epidermal basal precursors already possess a basal cell like identity immediately post fate segregation at 1 dpf dpf  we isolated itgb4+ basal precursors  krt4+ periderm cells from zebrafish embryos at 1 dpf  and adult itgb4+ basal cells from zebrafish caudal fin by fluorescence activated cell sorting. We then performed gene expression profiling analysis using data obtained from RNA seq of 3 different cells at 2 time points. Comparative gene expression profiling analysis of RNA seq data for embryonic basal cell precursors  embryonic periderm  and adult basal cells.", null, "pubmed:38265193", null, "Adult itgb4+ basal cells", "GSM7734879", null, "tissue:epidermal cells|cell type:adult fin epidermal basal cells|genotype:Tgitgb4:mcherry|geo loc name:missing|collection date:missing", "Adult itgb4+ basal cells", "High quality reads were mapped using hisat2 ver. 2.2.1 Trimmed sequence reads were mapped to GRCz11 using hisat2. Read count extraxction and normalization were performed using featureCounts ver. 2.0.3 Assembly: GRCz11 Supplementary files format and content: tab delimited text file includes basic gene information gene id  length  Genebank id  RNA type  gene name  product  raw count  RPKM and TPM values.", "epidermal cells", "Different types of cells were sorted by the fluorescence activated cell sorter SH800S SONY.", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer\u2019s protocol.", "Fish were maintained in a recirculating water system in a 14 h day/10 h night photoperiod at 28.5 \u00b0C. The wild type zebrafish Danio rerio strain used in this study was originally derived from the Tubingen strain and has been maintained in our facility for more than 10 years by inbreeding.", "cell type:adult fin epidermal basal cells|genotype:Tgitgb4:mcherry", "GSM7734879", "GSM7734879: Adult itgb4+ basal cells; Danio rerio; RNA Seq", "GSM7734879 r1", "GSM7734879", "1", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer's protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "DNBSEQ", "DNBSEQ-G400", null, "SRP457109", null, null, "Adult_itgb4mcherry_S230_R1_001.fastq.gz Adult_itgb4mcherry_S230_R2_001.fastq.gz", "fastq fastq", 5035430000.0, 12588575.0, "GSM7734879 r1", "0:200 1:200", "A:1339392170;C:1117195164;G:1254708952;T:1323174485;N:959229", 200, 200, null, null, 1339392170, 1117195164, 1254708952, 1323174485, 959229, "SRX21486772", "SRS18719072", "SRA1700438", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", 2, 0.01543, 0.02468, 0.00098, 0.00102, 0.99318, 0.99387, 0.47878, 0.31718, 200, 200, "T", "T", "mates < 9% mapping rate", "bgi", "bgi", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Japan", "2023-08-28", "Adult", "Adult", "Skin", "Surface Structure"], [25298, "SRR25764100", "SRX21486771", "SRS18719071", "SRP457109", "PRJNA1009810", "Live tracking of basal stem cells of the epidermis during growth  homeostasis  and injury response in zebrafish", "GSE241757", "Transcriptome Analysis", "Basal stem cells of the epidermis continuously differentiate into keratinocytes and replenish themselves via self renewal to maintain skin homeostasis. Numerous studies have attempted to reveal how basal cells undergo differentiation or self renewal; however  this has been hampered by a lack of robust basal cell markers and analytical platforms that allow single cell tracking. Here  we report integrin b4 itgb4 is a useful marker for basal cell labelling  irrespective of the body region  stage  and regenerative status. We employed Cre loxP recombination in combination with live cell tracking of single basal clones in the caudal fin and investigated the embryonic origin and behaviour of basal cells during fish growth and homeostasis. Although most basal cells  including those in fins  became quiescent in the adult stage  genetic cell ablation showed that basal cells were reactivated to either self renew or differentiate  depending on the injured cell type. Our findings provide a platform for quantitative in vivo imaging of basal stem cells applicable at wider stages and under various conditions and reveal unanticipated basal cell dynamism in response to distinct wound signals. Overall design: To investigate whether itgb4+ epidermal basal precursors already possess a basal cell like identity immediately post fate segregation at 1 dpf dpf  we isolated itgb4+ basal precursors  krt4+ periderm cells from zebrafish embryos at 1 dpf  and adult itgb4+ basal cells from zebrafish caudal fin by fluorescence activated cell sorting. We then performed gene expression profiling analysis using data obtained from RNA seq of 3 different cells at 2 time points. Comparative gene expression profiling analysis of RNA seq data for embryonic basal cell precursors  embryonic periderm  and adult basal cells.", null, "pubmed:38265193", null, "1 dpf itgb4+ embryonic basal precursors rep2", "GSM7734878", null, "tissue:epidermal cells|cell type:embryonic epidermal basal cells|genotype:Tgitgb4:mcherry|geo loc name:missing|collection date:missing", "1 dpf itgb4+ embryonic basal precursors rep2", "High quality reads were mapped using hisat2 ver. 2.2.1 Trimmed sequence reads were mapped to GRCz11 using hisat2. Read count extraxction and normalization were performed using featureCounts ver. 2.0.3 Assembly: GRCz11 Supplementary files format and content: tab delimited text file includes basic gene information gene id  length  Genebank id  RNA type  gene name  product  raw count  RPKM and TPM values.", "epidermal cells", "Different types of cells were sorted by the fluorescence activated cell sorter SH800S SONY.", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer\u2019s protocol.", "Fish were maintained in a recirculating water system in a 14 h day/10 h night photoperiod at 28.5 \u00b0C. The wild type zebrafish Danio rerio strain used in this study was originally derived from the Tubingen strain and has been maintained in our facility for more than 10 years by inbreeding.", "cell type:embryonic epidermal basal cells|genotype:Tgitgb4:mcherry", "GSM7734878", "GSM7734878: 1 dpf itgb4+ embryonic basal precursors rep2; Danio rerio; RNA Seq", "GSM7734878 r1", "GSM7734878", "1", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer's protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "DNBSEQ", "DNBSEQ-G400", null, "SRP457109", null, null, "itgb4mcherry_2_S229_R1_001.fastq.gz itgb4mcherry_2_S229_R2_001.fastq.gz", "fastq fastq", 6040245600.0, 15100614.0, "GSM7734878 r1", "0:200 1:200", "A:1588346322;C:1356558310;G:1524614760;T:1569583583;N:1142625", 200, 200, null, null, 1588346322, 1356558310, 1524614760, 1569583583, 1142625, "SRX21486771", "SRS18719071", "SRA1700438", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", 2, 0.0156, 0.0191, 0.00074, 0.00093, 0.99024, 0.99088, 0.47058, 0.47838, 200, 200, "T", "T", "mates < 9% mapping rate", "bgi", "bgi", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Japan", "2023-08-28", "Multi-stage", "Multi-stage", "Skin", "Surface Structure"], [25299, "SRR25764101", "SRX21486770", "SRS18719070", "SRP457109", "PRJNA1009810", "Live tracking of basal stem cells of the epidermis during growth  homeostasis  and injury response in zebrafish", "GSE241757", "Transcriptome Analysis", "Basal stem cells of the epidermis continuously differentiate into keratinocytes and replenish themselves via self renewal to maintain skin homeostasis. Numerous studies have attempted to reveal how basal cells undergo differentiation or self renewal; however  this has been hampered by a lack of robust basal cell markers and analytical platforms that allow single cell tracking. Here  we report integrin b4 itgb4 is a useful marker for basal cell labelling  irrespective of the body region  stage  and regenerative status. We employed Cre loxP recombination in combination with live cell tracking of single basal clones in the caudal fin and investigated the embryonic origin and behaviour of basal cells during fish growth and homeostasis. Although most basal cells  including those in fins  became quiescent in the adult stage  genetic cell ablation showed that basal cells were reactivated to either self renew or differentiate  depending on the injured cell type. Our findings provide a platform for quantitative in vivo imaging of basal stem cells applicable at wider stages and under various conditions and reveal unanticipated basal cell dynamism in response to distinct wound signals. Overall design: To investigate whether itgb4+ epidermal basal precursors already possess a basal cell like identity immediately post fate segregation at 1 dpf dpf  we isolated itgb4+ basal precursors  krt4+ periderm cells from zebrafish embryos at 1 dpf  and adult itgb4+ basal cells from zebrafish caudal fin by fluorescence activated cell sorting. We then performed gene expression profiling analysis using data obtained from RNA seq of 3 different cells at 2 time points. Comparative gene expression profiling analysis of RNA seq data for embryonic basal cell precursors  embryonic periderm  and adult basal cells.", null, "pubmed:38265193", null, "1 dpf itgb4+ embryonic basal precursors rep1", "GSM7734877", null, "tissue:epidermal cells|cell type:embryonic epidermal basal cells|genotype:Tgitgb4:mcherry|geo loc name:missing|collection date:missing", "1 dpf itgb4+ embryonic basal precursors rep1", "High quality reads were mapped using hisat2 ver. 2.2.1 Trimmed sequence reads were mapped to GRCz11 using hisat2. Read count extraxction and normalization were performed using featureCounts ver. 2.0.3 Assembly: GRCz11 Supplementary files format and content: tab delimited text file includes basic gene information gene id  length  Genebank id  RNA type  gene name  product  raw count  RPKM and TPM values.", "epidermal cells", "Different types of cells were sorted by the fluorescence activated cell sorter SH800S SONY.", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer\u2019s protocol.", "Fish were maintained in a recirculating water system in a 14 h day/10 h night photoperiod at 28.5 \u00b0C. The wild type zebrafish Danio rerio strain used in this study was originally derived from the Tubingen strain and has been maintained in our facility for more than 10 years by inbreeding.", "cell type:embryonic epidermal basal cells|genotype:Tgitgb4:mcherry", "GSM7734877", "GSM7734877: 1 dpf itgb4+ embryonic basal precursors rep1; Danio rerio; RNA Seq", "GSM7734877 r1", "GSM7734877", "1", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer's protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "DNBSEQ", "DNBSEQ-G400", null, "SRP457109", null, null, "itgb4mcherry_1_S20_R1_001.fastq.gz itgb4mcherry_1_S20_R2_001.fastq.gz", "fastq fastq", 7055215600.0, 17638039.0, "GSM7734877 r1", "0:200 1:200", "A:1894635584;C:1556854856;G:1769420212;T:1825520802;N:8784146", 200, 200, null, null, 1894635584, 1556854856, 1769420212, 1825520802, 8784146, "SRX21486770", "SRS18719070", "SRA1700438", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", 2, 0.01135, 0.01881, 0.00043, 0.00016, 0.9932, 0.99543, 0.46234, 0.43106, 200, 200, "T", "T", "mates < 9% mapping rate", "bgi", "bgi", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Japan", "2023-08-28", "Multi-stage", "Multi-stage", "Skin", "Surface Structure"], [25300, "SRR25764102", "SRX21486769", "SRS18719069", "SRP457109", "PRJNA1009810", "Live tracking of basal stem cells of the epidermis during growth  homeostasis  and injury response in zebrafish", "GSE241757", "Transcriptome Analysis", "Basal stem cells of the epidermis continuously differentiate into keratinocytes and replenish themselves via self renewal to maintain skin homeostasis. Numerous studies have attempted to reveal how basal cells undergo differentiation or self renewal; however  this has been hampered by a lack of robust basal cell markers and analytical platforms that allow single cell tracking. Here  we report integrin b4 itgb4 is a useful marker for basal cell labelling  irrespective of the body region  stage  and regenerative status. We employed Cre loxP recombination in combination with live cell tracking of single basal clones in the caudal fin and investigated the embryonic origin and behaviour of basal cells during fish growth and homeostasis. Although most basal cells  including those in fins  became quiescent in the adult stage  genetic cell ablation showed that basal cells were reactivated to either self renew or differentiate  depending on the injured cell type. Our findings provide a platform for quantitative in vivo imaging of basal stem cells applicable at wider stages and under various conditions and reveal unanticipated basal cell dynamism in response to distinct wound signals. Overall design: To investigate whether itgb4+ epidermal basal precursors already possess a basal cell like identity immediately post fate segregation at 1 dpf dpf  we isolated itgb4+ basal precursors  krt4+ periderm cells from zebrafish embryos at 1 dpf  and adult itgb4+ basal cells from zebrafish caudal fin by fluorescence activated cell sorting. We then performed gene expression profiling analysis using data obtained from RNA seq of 3 different cells at 2 time points. Comparative gene expression profiling analysis of RNA seq data for embryonic basal cell precursors  embryonic periderm  and adult basal cells.", null, "pubmed:38265193", null, "1 dpf krt4+ embryonic periderm cells rep2", "GSM7734876", null, "tissue:epidermal cells|cell type:embryonic periderm cells|genotype:Tgkrt4:creERt2;Olactb:loxP dsRed2 loxP egfp;cryaa:egfp|geo loc name:missing|collection date:missing", "1 dpf krt4+ embryonic periderm cells rep2", "High quality reads were mapped using hisat2 ver. 2.2.1 Trimmed sequence reads were mapped to GRCz11 using hisat2. Read count extraxction and normalization were performed using featureCounts ver. 2.0.3 Assembly: GRCz11 Supplementary files format and content: tab delimited text file includes basic gene information gene id  length  Genebank id  RNA type  gene name  product  raw count  RPKM and TPM values.", "epidermal cells", "Different types of cells were sorted by the fluorescence activated cell sorter SH800S SONY.", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer\u2019s protocol.", "Fish were maintained in a recirculating water system in a 14 h day/10 h night photoperiod at 28.5 \u00b0C. The wild type zebrafish Danio rerio strain used in this study was originally derived from the Tubingen strain and has been maintained in our facility for more than 10 years by inbreeding.", "cell type:embryonic periderm cells|genotype:Tgkrt4:creERt2;Olactb:loxP dsRed2 loxP egfp;cryaa:egfp", "GSM7734876", "GSM7734876: 1 dpf krt4+ embryonic periderm cells rep2; Danio rerio; RNA Seq", "GSM7734876 r1", "GSM7734876", "1", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer's protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "DNBSEQ", "DNBSEQ-G400", null, "SRP457109", null, null, "krt4cre-gfp_2_S4_R2_001.fastq.gz krt4cre-gfp_2_S4_R1_001.fastq.gz", "fastq fastq", 9439207600.0, 23598019.0, "GSM7734876 r1", "0:200 1:200", "A:2499982675;C:2086665360;G:2395002073;T:2454282285;N:3275207", 200, 200, null, null, 2499982675, 2086665360, 2395002073, 2454282285, 3275207, "SRX21486769", "SRS18719069", "SRA1700438", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", "Kawakami Lab, School of Life Science and Technology, Tokyo Institute of Technology", 2, 0.44931, 0.01877, 0.02236, 0.00115, 0.80012, 0.99032, 0.45473, 0.49453, 200, 200, "B", "T", "mate2 technical by mapping diff", "bgi", "bgi", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Japan", "2023-08-28", "Multi-stage", "Multi-stage", "Skin", "Surface Structure"], [25301, "SRR25764103", "SRX21486768", "SRS18719066", "SRP457109", "PRJNA1009810", "Live tracking of basal stem cells of the epidermis during growth  homeostasis  and injury response in zebrafish", "GSE241757", "Transcriptome Analysis", "Basal stem cells of the epidermis continuously differentiate into keratinocytes and replenish themselves via self renewal to maintain skin homeostasis. Numerous studies have attempted to reveal how basal cells undergo differentiation or self renewal; however  this has been hampered by a lack of robust basal cell markers and analytical platforms that allow single cell tracking. Here  we report integrin b4 itgb4 is a useful marker for basal cell labelling  irrespective of the body region  stage  and regenerative status. We employed Cre loxP recombination in combination with live cell tracking of single basal clones in the caudal fin and investigated the embryonic origin and behaviour of basal cells during fish growth and homeostasis. Although most basal cells  including those in fins  became quiescent in the adult stage  genetic cell ablation showed that basal cells were reactivated to either self renew or differentiate  depending on the injured cell type. Our findings provide a platform for quantitative in vivo imaging of basal stem cells applicable at wider stages and under various conditions and reveal unanticipated basal cell dynamism in response to distinct wound signals. Overall design: To investigate whether itgb4+ epidermal basal precursors already possess a basal cell like identity immediately post fate segregation at 1 dpf dpf  we isolated itgb4+ basal precursors  krt4+ periderm cells from zebrafish embryos at 1 dpf  and adult itgb4+ basal cells from zebrafish caudal fin by fluorescence activated cell sorting. We then performed gene expression profiling analysis using data obtained from RNA seq of 3 different cells at 2 time points. Comparative gene expression profiling analysis of RNA seq data for embryonic basal cell precursors  embryonic periderm  and adult basal cells.", null, "pubmed:38265193", null, "1 dpf krt4+ embryonic periderm cells rep1", "GSM7734875", null, "tissue:epidermal cells|cell type:embryonic periderm cells|genotype:Tgkrt4:creERt2;Olactb:loxP dsRed2 loxP egfp;cryaa:egfp|geo loc name:missing|collection date:missing", "1 dpf krt4+ embryonic periderm cells rep1", "High quality reads were mapped using hisat2 ver. 2.2.1 Trimmed sequence reads were mapped to GRCz11 using hisat2. Read count extraxction and normalization were performed using featureCounts ver. 2.0.3 Assembly: GRCz11 Supplementary files format and content: tab delimited text file includes basic gene information gene id  length  Genebank id  RNA type  gene name  product  raw count  RPKM and TPM values.", "epidermal cells", "Different types of cells were sorted by the fluorescence activated cell sorter SH800S SONY.", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer\u2019s protocol.", "Fish were maintained in a recirculating water system in a 14 h day/10 h night photoperiod at 28.5 \u00b0C. The wild type zebrafish Danio rerio strain used in this study was originally derived from the Tubingen strain and has been maintained in our facility for more than 10 years by inbreeding.", "cell type:embryonic periderm cells|genotype:Tgkrt4:creERt2;Olactb:loxP dsRed2 loxP egfp;cryaa:egfp", "GSM7734875", "GSM7734875: 1 dpf krt4+ embryonic periderm cells rep1; Danio rerio; RNA Seq", "GSM7734875 r1", "GSM7734875", "1", "The sorted cells were directly collected in a lysis buffer and used for RNA extraction using the RNeasy Plus Micro Kit Qiagen RNA libraries were prepared by MGIEasy\u2122 WES rapid library preparation kit according to the manufacturer's protocol.", null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "DNBSEQ", "DNBSEQ-G400", null, "SRP457109", null, null, "krt4cre-gfp_1_S19_R2_001.fastq.gz krt4cre-gfp_1_S19_R1_001.fastq.gz", "fastq fastq", 6119547200.0, 15298868.0, "GSM7734875 r1", "0:200 1:200", "A:1642059769;C:1349462427;G:1535937964;T:1584527048;N:7559992", 200, 200, 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Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. 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Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr28 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr28 5m 1", "zebrafish Nr28 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, "110307_DanioRerio_Nr28_5m_w1_s_2.fq", "fastq", 1004119144.0, 13212094.0, "zebrafish Nr28 5m 1 2", "0:76", "A:269579079;C:233472158;G:233160786;T:267709479;N:197642", 76, null, null, null, 269579079, 233472158, 233160786, 267709479, 197642, "SRX156096", "SRS346035", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92125, null, 0.09157, null, 0.72056, null, 0.4782, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2013-03-11", "Adult", "Adult", "Skin", "Surface Structure"], [36393, "SRR516131", "SRX156095", "SRS346034", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr27 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr27 5m 1", "zebrafish Nr27 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 1689987528.0, 22236678.0, "zebrafish Nr27 5m 1 1", "0:76", "A:462852366;C:385867681;G:380635625;T:460044585;N:587271", 76, null, null, null, 462852366, 385867681, 380635625, 460044585, 587271, "SRX156095", "SRS346034", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91288, null, 0.1015, null, 0.71293, null, 0.488, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36394, "SRR516136", "SRX156095", "SRS346034", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr27 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr27 5m 1", "zebrafish Nr27 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 421535824.0, 5546524.0, "zebrafish Nr27 5m 1 2", "0:76", "A:116434305;C:94958671;G:93876467;T:116039545;N:226836", 76, null, null, null, 116434305, 94958671, 93876467, 116039545, 226836, "SRX156095", "SRS346034", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.90776, null, 0.10459, null, 0.71399, null, 0.49081, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36395, "SRR516130", "SRX156094", "SRS346033", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr26 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr26 5m 1", "zebrafish Nr26 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 1633478944.0, 21493144.0, "zebrafish Nr26 5m 1 1", "0:76", "A:435251153;C:384938751;G:380973255;T:432171855;N:143930", 76, null, null, null, 435251153, 384938751, 380973255, 432171855, 143930, "SRX156094", "SRS346033", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92486, null, 0.08081, null, 0.71969, null, 0.50656, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36396, "SRR516129", "SRX156093", "SRS346032", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr25 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr25 5m 1", "zebrafish Nr25 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, "101021_DanioRerio_Nr25_5m_l7.fq", "fastq", 1893537644.0, 24914969.0, "zebrafish Nr25 5m 1 1", "0:76", "A:516246577;C:431589332;G:431233057;T:514385629;N:83049", 76, null, null, null, 516246577, 431589332, 431233057, 514385629, 83049, "SRX156093", "SRS346032", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.89513, null, 0.11188, null, 0.70591, null, 0.47827, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36397, "SRR516128", "SRX156092", "SRS346031", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr23 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr23 5m 1", "zebrafish Nr23 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, "101021_DanioRerio_Nr23_5m_l6.fq", "fastq", 2271040360.0, 29882110.0, "zebrafish Nr23 5m 1 1", "0:76", "A:602084142;C:529733757;G:524301688;T:614827528;N:93245", 76, null, null, null, 602084142, 529733757, 524301688, 614827528, 93245, "SRX156092", "SRS346031", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92194, null, 0.09655, null, 0.70463, null, 0.47059, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2013-03-11", "Adult", "Adult", "Skin", "Surface Structure"], [36398, "SRR516127", "SRX156091", "SRS346030", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr22 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr22 5m 1", "zebrafish Nr22 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2812847324.0, 37011149.0, "zebrafish Nr22 5m 1 1", "0:76", "A:760320146;C:645617435;G:643672544;T:763112485;N:124714", 76, null, null, null, 760320146, 645617435, 643672544, 763112485, 124714, "SRX156091", "SRS346030", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93272, null, 0.08997, null, 0.70755, null, 0.52272, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36399, "SRR516126", "SRX156090", "SRS346029", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 5 month. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  5m  skin  mRNA", "zebrafish Nr21 5m", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  5m  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr21 5m 1", "zebrafish Nr21 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2331404728.0, 30676378.0, "zebrafish Nr21 5m 1 1", "0:76", "A:630276643;C:539444103;G:529126097;T:632442560;N:115325", 76, null, null, null, 630276643, 539444103, 529126097, 632442560, 115325, "SRX156090", "SRS346029", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92747, null, 0.10341, null, 0.70414, null, 0.49626, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36400, "SRR516125", "SRX156089", "SRS346028", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 3.5 years. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  3.5y  skin  mRNA", "zebrafish Nr18 3.5y", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  3.5y  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr18 3.5y 1", "zebrafish Nr18 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2653618888.0, 34916038.0, "zebrafish Nr18 3.5y 1 1", "0:76", "A:728660211;C:598216653;G:601771842;T:724871715;N:98467", 76, null, null, null, 728660211, 598216653, 601771842, 724871715, 98467, "SRX156089", "SRS346028", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91227, null, 0.10895, null, 0.71802, null, 0.49946, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36401, "SRR516124", "SRX156088", "SRS346027", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 3.5 years. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  3.5y  skin  mRNA", "zebrafish Nr14 3.5y", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  3.5y  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr14 3.5y 1", "zebrafish Nr14 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2013201012.0, 26489487.0, "zebrafish Nr14 3.5y 1 1", "0:76", "A:554778161;C:456082219;G:445029202;T:557232699;N:78731", 76, null, null, null, 554778161, 456082219, 445029202, 557232699, 78731, "SRX156088", "SRS346027", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.90845, null, 0.1164, null, 0.71236, null, 0.48939, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36402, "SRR516123", "SRX156087", "SRS346026", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 3.5 years. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  3.5y  skin  mRNA", "zebrafish Nr13 3.5y", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  3.5y  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr13 3.5y 1", "zebrafish Nr13 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2680273532.0, 35266757.0, "zebrafish Nr13 3.5y 1 1", "0:76", "A:740111520;C:588329849;G:606909161;T:744802954;N:120048", 76, null, null, null, 740111520, 588329849, 606909161, 744802954, 120048, "SRX156087", "SRS346026", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.85899, null, 0.11949, null, 0.71595, null, 0.48655, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36403, "SRR516122", "SRX156086", "SRS346025", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 3.5 years. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  3.5y  skin  mRNA", "zebrafish Nr12 3.5y", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  3.5y  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr12 3.5y 1", "zebrafish Nr12 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2982635200.0, 39245200.0, "zebrafish Nr12 3.5y 1 1", "0:76", "A:829678392;C:650275344;G:659157904;T:842323646;N:1199914", 76, null, null, null, 829678392, 650275344, 659157904, 842323646, 1199914, "SRX156086", "SRS346025", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.88932, null, 0.12521, null, 0.71638, null, 0.53514, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36404, "SRR516121", "SRX156085", "SRS346024", "SRP013931", "PRJNA169239", "Initial analysis of transcript levels in zebrafish with advancing age", "PRJNA169239", "Other", "This project aims at an initial characterization of changes in gene expression in zebrafish with advancing age. Transcript levels are determined in several tissues of zebrafish with differing ages using RNA seq. Differentially expressed genes are determined to pinpoint genes that are differently regulated in young and old zebrafish. Results will be compared with other species to identify common pathways of ageing.", null, null, "Wild type zebrafish of the TueAB strain were kept in groups and only males were used for RNA isolation at an age of 3.5 years. Total RNA was isolated from skin using Trizol Invitrogen.", "1 male Danio rerio strain TueAB  3.5y  skin  mRNA", "zebrafish Nr11 3.5y", null, null, null, null, null, null, null, null, null, null, "1 male Danio rerio strain TueAB  3.5y  skin  mRNA  Illumina GA IIx  76 bp", "zebrafish Nr11 3.5y 1", "zebrafish Nr11 1", "1", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>76</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP013931", null, null, null, null, 2696915708.0, 35485733.0, "zebrafish Nr11 3.5y 1 1", "0:76", "A:749305835;C:599264137;G:607414252;T:740194221;N:737263", 76, null, null, null, 749305835, 599264137, 607414252, 740194221, 737263, "SRX156085", "SRS346024", "SRA054207", "Leibniz Institute for Age Research - Fritz Lipmann|Genome Analysis", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.86902, null, 0.11557, null, 0.71461, null, 0.49255, null, 76, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2012-06-26", "Adult", "Adult", "Skin", "Surface Structure"], [36712, "SRR835167", "SRX271963", "SRS416259", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 116h 53", "GSM1129625", null, "tissue:melanocytes|hpf", "dm 116h 53", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129625", "GSM1129625: dm 116h 53; Danio rerio; RNA Seq", "GSM1129625 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129625", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_116h_53_440.fq.bz2", "fastq", 203842506.0, 4853393.0, "GSM1129625 r1", "0:42", "A:49784918;C:48871504;G:45835011;T:59338271;N:12802", 42, null, null, null, 49784918, 48871504, 45835011, 59338271, 12802, "SRX271963", "SRS416259", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.82741, null, 0.14973, null, 0.84893, null, 0.53644, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36713, "SRR835166", "SRX271962", "SRS416258", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 97h 41 440", "GSM1129624", null, "tissue:melanocytes|hpf", "dm 97h 41 440", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129624", "GSM1129624: dm 97h 41 440; Danio rerio; RNA Seq", "GSM1129624 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129624", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_97h_41_440.fq.bz2", "fastq", 199638768.0, 4753304.0, "GSM1129624 r1", "0:42", "A:45958194;C:50367228;G:48451580;T:54849238;N:12528", 42, null, null, null, 45958194, 50367228, 48451580, 54849238, 12528, "SRX271962", "SRS416258", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.85741, null, 0.09897, null, 0.84112, null, 0.53299, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36714, "SRR835165", "SRX271961", "SRS416257", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h FM 351", "GSM1129623", null, "tissue:melanocytes|hpf", "dm 77h FM 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129623", "GSM1129623: dm 77h FM 351; Danio rerio; RNA Seq", "GSM1129623 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129623", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_FM_351_ATT.fq.bz2", "fastq", 194385384.0, 5399594.0, "GSM1129623 r1", "0:36", "A:45805148;C:47931006;G:41165284;T:59482433;N:1513", 36, null, null, null, 45805148, 47931006, 41165284, 59482433, 1513, "SRX271961", "SRS416257", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.77748, null, 0.07764, null, 0.87687, null, 0.51575, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36715, "SRR835164", "SRX271960", "SRS416256", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 52 440", "GSM1129622", null, "tissue:melanocytes|hpf", "dm 77h 52 440", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129622", "GSM1129622: dm 77h 52 440; Danio rerio; RNA Seq", "GSM1129622 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129622", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_52_440.fq.bz2", "fastq", 105047586.0, 2501133.0, "GSM1129622 r1", "0:42", "A:24711000;C:26459041;G:24869198;T:29001957;N:6390", 42, null, null, null, 24711000, 26459041, 24869198, 29001957, 6390, "SRX271960", "SRS416256", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.88862, null, 0.09219, null, 0.83611, null, 0.55476, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36716, "SRR835163", "SRX271959", "SRS416255", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 43 440", "GSM1129621", null, "tissue:melanocytes|hpf", "dm 77h 43 440", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129621", "GSM1129621: dm 77h 43 440; Danio rerio; RNA Seq", "GSM1129621 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129621", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_43_440.fq.bz2", "fastq", 371553084.0, 8846502.0, "GSM1129621 r1", "0:42", "A:84994407;C:91269417;G:87188701;T:108077735;N:22824", 42, null, null, null, 84994407, 91269417, 87188701, 108077735, 22824, "SRX271959", "SRS416255", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.86495, null, 0.09748, null, 0.83684, null, 0.52833, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36717, "SRR835162", "SRX271958", "SRS416254", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 28 399s62", "GSM1129620", null, "tissue:melanocytes|hpf", "dm 77h 28 399s62", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129620", "GSM1129620: dm 77h 28 399s62; Danio rerio; RNA Seq", "GSM1129620 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129620", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, null, null, 731276158.0, 3620179.0, "GSM1129620 r1", "0:101 1:101", "A:173989639;C:199270484;G:188281318;T:169725601;N:9116", 101, 101, null, null, 173989639, 199270484, 188281318, 169725601, 9116, "SRX271958", "SRS416254", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 2, 0.84122, 0.79964, 0.02274, 0.02168, 0.89645, 0.89073, 0.63341, 0.63015, 101, 101, "B", "B", "biological fallback assumption", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36718, "SRR835161", "SRX271957", "SRS416253", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 28 399s61", "GSM1129619", null, "tissue:melanocytes|hpf", "dm 77h 28 399s61", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129619", "GSM1129619: dm 77h 28 399s61; Danio rerio; RNA Seq", "GSM1129619 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129619", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_28_399s61_CACT.fq.bz2 dm_77h_28_399s62_CACT.fq.bz2", "fastq fastq", 731276158.0, 3620179.0, "GSM1129619 r1", "0:101 1:101", "A:173989639;C:199270484;G:188281318;T:169725601;N:9116", 101, 101, null, null, 173989639, 199270484, 188281318, 169725601, 9116, "SRX271957", "SRS416253", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 2, 0.84121, 0.79968, 0.02287, 0.02141, 0.89658, 0.89051, 0.63318, 0.63061, 101, 101, "B", "B", "biological fallback assumption", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36719, "SRR835160", "SRX271956", "SRS416252", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 28 436", "GSM1129618", null, "tissue:melanocytes|hpf", "dm 77h 28 436", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129618", "GSM1129618: dm 77h 28 436; Danio rerio; RNA Seq", "GSM1129618 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129618", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_28_436_CACT.fq.bz2", "fastq", 214752678.0, 5113159.0, "GSM1129618 r1", "0:42", "A:50210098;C:62230664;G:49216487;T:52926929;N:168500", 42, null, null, null, 50210098, 62230664, 49216487, 52926929, 168500, "SRX271956", "SRS416252", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.80956, null, 0.02412, null, 0.87024, null, 0.62475, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36720, "SRR835159", "SRX271955", "SRS416251", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 28 351", "GSM1129617", null, "tissue:melanocytes|hpf", "dm 77h 28 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129617", "GSM1129617: dm 77h 28 351; Danio rerio; RNA Seq", "GSM1129617 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129617", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_28_351_CACT.fq.bz2", "fastq", 84584556.0, 2349571.0, "GSM1129617 r1", "0:36", "A:19843286;C:24761303;G:18855233;T:21124051;N:683", 36, null, null, null, 19843286, 24761303, 18855233, 21124051, 683, "SRX271955", "SRS416251", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.75246, null, 0.02663, null, 0.87054, null, 0.59024, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36721, "SRR835158", "SRX271954", "SRS416250", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 27 436", "GSM1129616", null, "tissue:melanocytes|hpf", "dm 77h 27 436", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129616", "GSM1129616: dm 77h 27 436; Danio rerio; RNA Seq", "GSM1129616 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129616", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_27_436_TTAT.fq.bz2", "fastq", 194114466.0, 4621773.0, "GSM1129616 r1", "0:42", "A:44579558;C:47759429;G:44539735;T:57080409;N:155335", 42, null, null, null, 44579558, 47759429, 44539735, 57080409, 155335, "SRX271954", "SRS416250", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.80787, null, 0.04019, null, 0.86263, null, 0.57373, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36722, "SRR835157", "SRX271953", "SRS416249", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 27 351", "GSM1129615", null, "tissue:melanocytes|hpf", "dm 77h 27 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129615", "GSM1129615: dm 77h 27 351; Danio rerio; RNA Seq", "GSM1129615 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129615", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_27_351_TTAT.fq.bz2", "fastq", 113087304.0, 3141314.0, "GSM1129615 r1", "0:36", "A:26171623;C:27323878;G:24871328;T:34719586;N:889", 36, null, null, null, 26171623, 27323878, 24871328, 34719586, 889, "SRX271953", "SRS416249", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.75375, null, 0.04337, null, 0.85878, null, 0.5311, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36723, "SRR835156", "SRX271952", "SRS416248", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 26 433", "GSM1129614", null, "tissue:melanocytes|hpf", "dm 77h 26 433", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129614", "GSM1129614: dm 77h 26 433; Danio rerio; RNA Seq", "GSM1129614 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129614", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_26_433_AGAT.fq.bz2", "fastq", 34005804.0, 809662.0, "GSM1129614 r1", "0:42", "A:8766789;C:8066807;G:8338043;T:8833768;N:397", 42, null, null, null, 8766789, 8066807, 8338043, 8833768, 397, "SRX271952", "SRS416248", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.81802, null, 0.05684, null, 0.88787, null, 0.56756, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36724, "SRR835155", "SRX271951", "SRS416247", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 26 351", "GSM1129613", null, "tissue:melanocytes|hpf", "dm 77h 26 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129613", "GSM1129613: dm 77h 26 351; Danio rerio; RNA Seq", "GSM1129613 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129613", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_26_351_AGAT.fq.bz2", "fastq", 101340432.0, 2815012.0, "GSM1129613 r1", "0:36", "A:26592120;C:23518331;G:24503636;T:26725495;N:850", 36, null, null, null, 26592120, 23518331, 24503636, 26725495, 850, "SRX271951", "SRS416247", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.75158, null, 0.05787, null, 0.88986, null, 0.57167, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36725, "SRR835154", "SRX271950", "SRS416245", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 25 399s62", "GSM1129612", null, "tissue:melanocytes|hpf", "dm 77h 25 399s62", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129612", "GSM1129612: dm 77h 25 399s62; Danio rerio; RNA Seq", "GSM1129612 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129612", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, null, null, 1715809008.0, 8494104.0, "GSM1129612 r1", "0:101 1:101", "A:405310561;C:444474389;G:467897835;T:398105078;N:21145", 101, 101, null, null, 405310561, 444474389, 467897835, 398105078, 21145, "SRX271950", "SRS416245", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 2, 0.66327, 0.58026, 0.07911, 0.07288, 0.91721, 0.9083, 0.56907, 0.5363, 101, 101, "B", "B", "biological fallback assumption", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36726, "SRR835153", "SRX271949", "SRS416246", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 25 399s61", "GSM1129611", null, "tissue:melanocytes|hpf", "dm 77h 25 399s61", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129611", "GSM1129611: dm 77h 25 399s61; Danio rerio; RNA Seq", "GSM1129611 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129611", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "PAIRED", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_25_399s61_TCTT.fq.bz2 dm_77h_25_399s62_TCTT.fq.bz2", "fastq fastq", 1715809008.0, 8494104.0, "GSM1129611 r1", "0:101 1:101", "A:405310561;C:444474389;G:467897835;T:398105078;N:21145", 101, 101, null, null, 405310561, 444474389, 467897835, 398105078, 21145, "SRX271949", "SRS416246", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 2, 0.66325, 0.58027, 0.0792, 0.07327, 0.91741, 0.90847, 0.56374, 0.53632, 101, 101, "B", "B", "biological fallback assumption", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36727, "SRR835152", "SRX271948", "SRS416244", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 25 433", "GSM1129610", null, "tissue:melanocytes|hpf", "dm 77h 25 433", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129610", "GSM1129610: dm 77h 25 433; Danio rerio; RNA Seq", "GSM1129610 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129610", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_25_433_TCTT.fq.bz2", "fastq", 489386604.0, 11652062.0, "GSM1129610 r1", "0:42", "A:99597908;C:134563195;G:119005661;T:136214210;N:5630", 42, null, null, null, 99597908, 134563195, 119005661, 136214210, 5630, "SRX271948", "SRS416244", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.79072, null, 0.08396, null, 0.87497, null, 0.51318, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36728, "SRR835151", "SRX271947", "SRS416243", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 77h 25 351", "GSM1129609", null, "tissue:melanocytes|hpf", "dm 77h 25 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129609", "GSM1129609: dm 77h 25 351; Danio rerio; RNA Seq", "GSM1129609 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129609", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_77h_25_351_TCTT.fq.bz2", "fastq", 98765532.0, 2743487.0, "GSM1129609 r1", "0:36", "A:19387910;C:27338385;G:23220852;T:28817575;N:810", 36, null, null, null, 19387910, 27338385, 23220852, 28817575, 810, "SRX271947", "SRS416243", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.75369, null, 0.08166, null, 0.87667, null, 0.51755, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36729, "SRR835150", "SRX271946", "SRS416242", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 58h 45 440", "GSM1129608", null, "tissue:melanocytes|hpf", "dm 58h 45 440", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129608", "GSM1129608: dm 58h 45 440; Danio rerio; RNA Seq", "GSM1129608 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129608", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_58h_45_440.fq.bz2", "fastq", 203660982.0, 4849071.0, "GSM1129608 r1", "0:42", "A:46576136;C:50204145;G:47522343;T:59345685;N:12673", 42, null, null, null, 46576136, 50204145, 47522343, 59345685, 12673, "SRX271946", "SRS416242", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.86133, null, 0.0978, null, 0.84565, null, 0.53118, null, 42, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [36730, "SRR835149", "SRX271945", "SRS416241", "SRP021517", "PRJNA198908", "Gene Expression Analysis of Zebrafish Melanocytes  Iridophores  and Retinal Pigmented Epithelium Reveals Indicators of Biological Function and Developmental Origin", "GSE46387", "Transcriptome Analysis", "In order to facilitate understanding of pigment cell biology  we developed a method to concomitantly purify melanocytes  iridophores  and retinal pigmented epithelium from zebrafish  and analyzed their transcriptomes.  Comparing expression data from these cell types and whole embryos allowed us to reveal gene expression co enrichment in melanocytes and retinal pigmented epithelium  as well as in melanocytes and iridophores.  We found 214 genes co enriched in melanocytes and retinal pigmented epithelium  indicating the shared functions of melanin producing cells.  We found 62 genes significantly co enriched in melanocytes and iridophores  illustrative of their shared developmental origins from the neural crest.  This is also the first analysis of the iridophore transcriptome.  Gene expression analysis for iridophores revealed extensive enrichment of specific enzymes to coordinate production of their guanine based reflective pigment.  We speculate the coordinated upregulation of specific enzymes from several metabolic pathways recycles the rate limiting substrate for purine synthesis  phosphoribosyl pyrophosphate  thus constituting a guanine cycle.  The purification procedure and expression analysis described here  along with the accompanying transcriptome wide expression data  provide the first mRNA sequencing data for multiple purified zebrafish pigment cell types  and will be a useful resource for further studies of pigment cell biology. Overall design: mRNA profiles of zebrafish pigment cells were generated using Illumina GAIIX sequencing", null, "pubmed:23874447", null, "dm 58h 23 351", "GSM1129607", null, "tissue:melanocytes|hpf", "dm 58h 23 351", "Align sequences using Novoalign to cDNA database Calculate RPKMs from Novoalign output using Perl Genome build: Non redundant database of 25 102 cDNAs generated by Higdon et al 2013  based on NCBI's RefSeq build from 8/29/2012  current RefSeq available at ftp://ftp.ncbi.nlm.nih.gov/refseq/D rerio/mRNA Prot/zebrafish.rna.fna.gz Supplementary files format and content: Excell file containing RPKMs of each library", "melanocytes", null, "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", null, "hpf", "GSM1129607", "GSM1129607: dm 58h 23 351; Danio rerio; RNA Seq", "GSM1129607 1", null, "1", "This study was carried out in accordance with the Washington University Animal Use Committee guidelines under approved protocol #20110236.  Zebrafish were reared and bred according to standard protocols.  The fish used in this study were homozygous for a temperature sensitive allele of micropthalmia transcription factor  mitfavc7.  This mutant facilitated the collection of RPE  as mitfa is not required for RPE development in zebrafish.  Melanocytes  iridophores  and RPE develop normally at 25\u00b0C in mitfavc7.  When held at 32\u00b0C  the neural crest derived melanocytes do not develop  but RPE and iridophores develop normally.  All melanocyte and iridophore samples were incubated at 25\u00b0C prior to collection.  Fish were anesthetized with Tricaine  rinsed with Ca   Mg  DPBS Sigma  D8537  and immersed in 100mL TrypLE Express Invitrogen  12604039 per 1000 fish.  Fish were incubated at 37\u00b0C and shaken at 100rpm for 15 20 minutes  followed by trituration with a Pasteur pipette to remove eyes from larva.  post separation of eyes and larva  each group was placed in TrypLE Express and shaken at 100rpm at 37\u00b0C for 1 1.5 hr.  Dissociated cells were filtered through a 120uM screen into 50 mL tubes.  Remaining intact tissue was triturated 10 20 times  and again filtered through a 120uM screen into the dissociated cells.  Dissociated cells were pelleted in a swinging bucket rotor Eppendorf 5810 R at 500 relative centrifugal force rcf for 5 minutes at 4\u00b0C  then resuspended in 1mL cold isotonic Percoll Sigma  P1644 by gentle pipetting.  Isotonic Percoll was prepared by mixing 1 part 10X PBS with 9 parts Percoll.  Resuspended cells were transferred to 1.6mL Eppendorf tubes and spun at 2000rcf for 5 minutes at 4\u00b0C in a swinging bucket rotor for isopycnic separation.  Pigment cells in the pellet were then resuspended in 400\u00b5L of ice cold DPBS with 2% fetal calf serum FCS  and placed onto preformed Percoll density gradients.  Preformed gradients were prepared via centrifugation of 1mL aliquots of isotonic Percoll in 1.6mL tubes at 10 000rcf for 15 minutes at 4\u00b0C in a fixed angle micro centrifuge Eppendorf 5415 R.  Tubes containing preformed Percoll gradients with overlying cell suspensions were centrifuged in a swinging bucket rotor at 2000rcf for 10 minutes at 4\u00b0C.  Following centrifugation  overlying Percoll was aspirated  leaving the final 100\u00b5L containing the pigment cell pellet.  Cells were resuspended with 50\u00b5L of cold DPBS with 2% FCS and transferred to a clean 1.6mL tube containing 500\u00b5L of cold DPBS with 2% FCS  and kept on ice until mRNA extraction or FACS.      FACS   We used the inherent properties of the pigmented cells to perform Fluorescence Activated Cell Sorting FACS.  Following resuspension in 500\u00b5L cold DPBS with 2% FCS  the enriched cell populations were screened through a 30uM cell filter Partec  04 0042 2316.  Cells were analyzed and sorted with a Dako MoFlo cell sorter using a 120uM nozzle at a drop drive DD frequency of 22390Hz.  Cells were illuminated using a 488 nm laser.  Cells were gated on two attributes to separate cells from each other and from cellular debris.  Cellular debris was detected using forward and side scatter  selecting against the smallest particles 1\u00b5m or less.  Cells were sorted based on detection using 510 530nm and 575 595nm filters  corresponding to FL1 and FL2 in Figure 1D  respectively.  When excited by the 488 nm laser  the autofluorescence of iridophores is clearly detectable in these channels as a group of cells extending at a 45 degree line in the upper right quadrant.  Melanocytes and RPE do not autofluoresce with this intensity when excited by the 488nm laser  and cluster at the lower left of the FACS plot.  Cells were collected into ice cold DPBS with 2% FCS and kept on ice until mRNA extraction. Pigment cell cDNA library construction was as follows.  For mRNA extraction the Dynabeads\u00ae mRNA DIRECT Kit Invitrogen was used per manufacturer's instructions.  Following mRNA elution from the Dynabeads  first strand cDNA synthesis was performed using MMLV reverse transcriptase Clontech using an anchored polyT primer tailed with a universal primer sequence See Table S3 for primer sequences and Figure 2 for pigment cell cDNA library construction overview.  A universal primer sequence was also added to the three prime end of the first strand by template switching  allowing for PCR amplification of the resultant cDNA  [43 44].  Following PCR amplification using the high fidelity polymerase LA Taq TaKaRa   PCR cycle: 95C for 1 minute  followed by 20 cycles of 98C for 25 seconds  60C for 1 minute  68C for 20 minutes  cDNA was digested with AluI and RsaI restriction enzymes NEB.  Blunt end enzymatic fragmentation of cDNA was used instead of sonication and gel extraction to minimize loss of sample material and eliminate the end repair step of Illumina library preparation.  Since this reduced representation strategy might miss short cDNAs that lack both restriction sites  we sought to avoid this by including enzyme recognition sites within the cDNA amplification primers.  This allows for the inclusion of short cDNAs in our libraries.  Standard Illumina library preparations followed  performed by the Genome Technology Access Center GTAC at Washington University in St. Louis http://gtac.wustl.edu.  In brief  a single A was added to the 3\u2019 end of each strand  Y adapters ligated  and library enrichment PCR performed  followed by gel extraction size selection for fragments ranging from 200 400 base pairs in length.  Illumina library construction of pooled 3dpf embryos was performed by GTAC from total RNA extracted with Trizol reagent as previously described [45].  No PCR amplification of whole embryo cDNA was performed prior to Illumina adapter ligation and library enrichment.  Sequencing was performed on the GAIIX Illumina platform.", "GEO Accession:GSM1129607", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP021517", null, null, "dm_58h_23_351_GAAT.fq.bz2", "fastq", 106123464.0, 2947874.0, "GSM1129607 r1", "0:36", "A:26495114;C:26882652;G:25518440;T:27226454;N:804", 36, null, null, null, 26495114, 26882652, 25518440, 27226454, 804, "SRX271945", "SRS416241", "SRA074390", "GEO", "Rob Mitra, Genetics, Washington University", 1, 0.78561, null, 0.06331, null, 0.85107, null, 0.46073, null, 36, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "3prime", "poly_a", "unknown", "bulk", "unknown", "unknown", null, "United States", "2013-04-25", "Larval", "Larval", "Skin", "Surface Structure"], [38116, "SRR1551799", "SRX681419", "SRS685080", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "keratocytes from 4dpf embryos", "keratocytes from 4dpf embryos", null, "breed:TL|strain:TL|age:4dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:4dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "transcriptome from 4dpf keratocytes  replicate 3", "4dpf 3", "4dpf 3", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>40</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane4_4dpf707.fastq", "fastq", 1001179200.0, 25029480.0, "4dpf 3", "0:40", "A:253770714;C:242097985;G:244642013;T:260557997;N:110491", 40, null, null, null, 253770714, 242097985, 244642013, 260557997, 110491, "SRX681419", "SRS685080", "SRA179326", "Stanford University|Julie Theriot", "Stanford University", 1, 0.90546, null, 0.2101, null, 0.75185, null, 0.49242, null, 40, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2015-04-07", "Larval", "Larval", "Skin", "Surface Structure"], [38117, "SRR1551798", "SRX681418", "SRS685080", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "keratocytes from 4dpf embryos", "keratocytes from 4dpf embryos", null, "breed:TL|strain:TL|age:4dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:4dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "Transcriptome from 4dpf keratocytes  replicate 2", "4dpf 2", "4dpf 2", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>40</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane7_4col.fastq", "fastq", 1700157430.0, 44740985.0, "4dpf 2", "0:38", "A:394789834;C:418821057;G:504852317;T:369111176;N:12583046", 38, null, null, null, 394789834, 418821057, 504852317, 369111176, 12583046, "SRX681418", "SRS685080", "SRA179326", "Stanford University|Julie Theriot", "Stanford University", 1, 0.68441, null, 0.12529, null, 0.8338, null, 0.45494, null, 38, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2014-08-14", "Larval", "Larval", "Skin", "Surface Structure"], [38118, "SRR1551797", "SRX681417", "SRS685080", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "keratocytes from 4dpf embryos", "keratocytes from 4dpf embryos", null, "breed:TL|strain:TL|age:4dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:4dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "4dpf keratocyte transcriptome  replicate 1", "4dpf 1", "4dpf 1", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>47</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane7_1_4dpf.fastq", "fastq", 762138229.0, 16215707.0, "4dpf 1", "0:47", "A:183899711;C:180347476;G:190086042;T:189460942;N:18344058", 47, null, null, null, 183899711, 180347476, 190086042, 189460942, 18344058, "SRX681417", "SRS685080", "SRA179326", "Stanford University|Julie Theriot", "Stanford University", 1, 0.85884, null, 0.2135, null, 0.80077, null, 0.50539, null, 47, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2014-08-15", "Larval", "Larval", "Skin", "Surface Structure"], [38119, "SRR1551796", "SRX681416", "SRS685079", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "Transcriptome from 2dpf keratocytes", "keratocytes from 2dpf embryos", null, "breed:TL|strain:TL|age:2dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:2dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "Transcriptome from 2dpf keratocytes  replicate 3", "2dpf 3", "2dpf 3", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>40</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane3_2dpf629.fastq", "fastq", 551787520.0, 13794688.0, "2dpf 3", "0:40", "A:138660463;C:132605577;G:134640312;T:145852714;N:28454", 40, null, null, null, 138660463, 132605577, 134640312, 145852714, 28454, "SRX681416", "SRS685079", "SRA179323", "Stanford University|Julie Theriot", "Stanford University", 1, 0.91375, null, 0.21598, null, 0.75317, null, 0.47881, null, 40, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2014-08-14", "Hatching", "Embryo", "Skin", "Surface Structure"], [38120, "SRR1551795", "SRX681415", "SRS685079", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "Transcriptome from 2dpf keratocytes", "keratocytes from 2dpf embryos", null, "breed:TL|strain:TL|age:2dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:2dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "Transcriptome from 2dpf keratocytes", "2dpf 2", "2dpf 2", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>40</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane1_2dpf.fastq", "fastq", 796874112.0, 22135392.0, "2dpf 2", "0:36", "A:209956517;C:184659684;G:185142987;T:216055902;N:1059022", 36, null, null, null, 209956517, 184659684, 185142987, 216055902, 1059022, "SRX681415", "SRS685079", "SRA179323", "Stanford University|Julie Theriot", "Stanford University", 1, 0.89178, null, 0.17837, null, 0.7609, null, 0.49774, null, 36, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2015-04-07", "Hatching", "Embryo", "Skin", "Surface Structure"], [38121, "SRR1551782", "SRX681402", "SRS685079", "SRP045504", "PRJNA258223", "Danio rerio strain:TL Transcriptome or Gene expression", "PRJNA258223", "Other", "Transcriptome profiling of keratocytes derived from 2dpf and 4dpf embryos", null, null, null, "Transcriptome from 2dpf keratocytes", "keratocytes from 2dpf embryos", null, "breed:TL|strain:TL|age:2dpf|biomaterial provider:Julie Theriot  Stanford University|sex:not collected|tissue:keratocytes skin|cell type:keratocyte|dev stage:2dpf|BioSampleModel:Model organism or animal", null, null, null, null, null, null, null, null, "2dpf keratocytes  replicate 1", "2dpf 1", "2dpf 1", "RNA was extracted from approximately 1 x 105 keratocytes per sample using Trizol Invitrogen.  Three biological replicates were collected for each developmental stage.  Ribosomal RNA was depleted using the Ribo Zero magnetic kit Epicentre.  The remaining mRNA was then fragmented with 8 minutes incubation in 50mM sodium carbonate/bicarbonate  1mM EDTA  pH 9.2 at 95 degrees.  To prepare cDNA  first strand synthesis was performed with Superscript III Invitrogen using random hexamer priming  and second strand synthesis was performed with DNA Polymerase I NEB.  Illumina libraries were prepared from the cDNA in an automated fashion using the Illumina TruSeq sample prep kit with TruSeq adapters on a SPRIworks System I Beckman Coulter.  Sequencing reactions were performed on an Illumina Genome Analyzer IIX according to manufacturer\u2019s instructions to generate 40nt single ended reads.", null, null, "RNA-Seq", "TRANSCRIPTOMIC", "PolyA", "SINGLE", "ILLUMINA", "Illumina Genome Analyzer IIx", "<SPOT_DESCRIPTOR><SPOT_DECODE_SPEC><SPOT_LENGTH>40</SPOT_LENGTH><READ_SPEC><READ_INDEX>0</READ_INDEX><READ_CLASS>Application Read</READ_CLASS><READ_TYPE>Forward</READ_TYPE><BASE_COORD>1</BASE_COORD></READ_SPEC></SPOT_DECODE_SPEC></SPOT_DESCRIPTOR>", "SRP045504", null, null, "lane6_2dpf.fastq", "fastq", 647944118.0, 17051161.0, "2dpf 1", "0:38", "A:145735255;C:167786338;G:186677806;T:141598169;N:6146550", 38, null, null, null, 145735255, 167786338, 186677806, 141598169, 6146550, "SRX681402", "SRS685079", "SRA179323", "Stanford University|Julie Theriot", "Stanford University", 1, 0.7695, null, 0.15816, null, 0.81491, null, 0.44972, null, 38, null, "B", null, "usable mapping rate", "illumina", "early_illumina", "unknown", "poly_a", "trueseq", "bulk", "unknown", "unknown", null, "United States", "2014-08-14", "Hatching", "Embryo", "Skin", "Surface Structure"], [38389, "SRR1821850", "SRX893451", "SRS859516", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin208 36m 3.75nM roten1 8w rep6", "GSM1620997", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin208 36m 3.75nM roten1 8w rep6", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620997", "GSM1620997: skin208 36m 3.75nM roten1 8w rep6; Danio rerio; RNA Seq", "GSM1620997", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620997", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "208_skin.fastq.gz", "fastq", 1960697700.0, 39213954.0, "GSM1620997 r1", "0:50", "A:532792887;C:452528560;G:441240826;T:533522462;N:612965", 50, null, null, null, 532792887, 452528560, 441240826, 533522462, 612965, "SRX893451", "SRS859516", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92216, null, 0.09326, null, 0.72147, null, 0.49795, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38390, "SRR1821849", "SRX893450", "SRS859517", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin207 36m 3.75nM roten1 8w rep5", "GSM1620996", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin207 36m 3.75nM roten1 8w rep5", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620996", "GSM1620996: skin207 36m 3.75nM roten1 8w rep5; Danio rerio; RNA Seq", "GSM1620996", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620996", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "207_skin.fastq.gz", "fastq", 1644999850.0, 32899997.0, "GSM1620996 r1", "0:50", "A:444439104;C:381379643;G:371782441;T:447032625;N:366037", 50, null, null, null, 444439104, 381379643, 371782441, 447032625, 366037, "SRX893450", "SRS859517", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.90733, null, 0.0971, null, 0.71518, null, 0.48175, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38391, "SRR1821848", "SRX893449", "SRS859518", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin206 36m 3.75nM roten1 8w rep4", "GSM1620995", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin206 36m 3.75nM roten1 8w rep4", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620995", "GSM1620995: skin206 36m 3.75nM roten1 8w rep4; Danio rerio; RNA Seq", "GSM1620995", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620995", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "206_skin.fastq.gz", "fastq", 2333175800.0, 46663516.0, "GSM1620995 r1", "0:50", "A:629018600;C:541489412;G:532198591;T:629985712;N:483485", 50, null, null, null, 629018600, 541489412, 532198591, 629985712, 483485, "SRX893449", "SRS859518", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91718, null, 0.10454, null, 0.70974, null, 0.47753, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38392, "SRR1821847", "SRX893448", "SRS859519", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin205 36m 3.75nM roten1 8w rep3", "GSM1620994", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin205 36m 3.75nM roten1 8w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620994", "GSM1620994: skin205 36m 3.75nM roten1 8w rep3; Danio rerio; RNA Seq", "GSM1620994", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620994", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "205_skin.fastq.gz", "fastq", 1885120700.0, 37702414.0, "GSM1620994 r1", "0:50", "A:509247143;C:431669478;G:422352918;T:511569209;N:10281952", 50, null, null, null, 509247143, 431669478, 422352918, 511569209, 10281952, "SRX893448", "SRS859519", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.90066, null, 0.107, null, 0.71415, null, 0.48729, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38393, "SRR1821846", "SRX893447", "SRS859520", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin204 36m 3.75nM roten1 8w rep2", "GSM1620993", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin204 36m 3.75nM roten1 8w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620993", "GSM1620993: skin204 36m 3.75nM roten1 8w rep2; Danio rerio; RNA Seq", "GSM1620993", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620993", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "204_skin.fastq.gz", "fastq", 2039214400.0, 40784288.0, "GSM1620993 r1", "0:50", "A:554630438;C:468936669;G:460030131;T:555276071;N:341091", 50, null, null, null, 554630438, 468936669, 460030131, 555276071, 341091, "SRX893447", "SRS859520", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91261, null, 0.11181, null, 0.70832, null, 0.4889, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38394, "SRR1821845", "SRX893446", "SRS859521", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin203 36m 3.75nM roten1 8w rep1", "GSM1620992", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "skin203 36m 3.75nM roten1 8w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:8 weeks", "GSM1620992", "GSM1620992: skin203 36m 3.75nM roten1 8w rep1; Danio rerio; RNA Seq", "GSM1620992", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620992", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "203_skin.fastq.gz", "fastq", 2169512850.0, 43390257.0, "GSM1620992 r1", "0:50", "A:583644936;C:503133232;G:492417551;T:585152349;N:5164782", 50, null, null, null, 583644936, 503133232, 492417551, 585152349, 5164782, "SRX893446", "SRS859521", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91263, null, 0.1004, null, 0.71376, null, 0.48566, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38395, "SRR1821844", "SRX893445", "SRS859522", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin172 36m 3.75nM roten1 3w rep5", "GSM1620991", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "skin172 36m 3.75nM roten1 3w rep5", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "GSM1620991", "GSM1620991: skin172 36m 3.75nM roten1 3w rep5; Danio rerio; RNA Seq", "GSM1620991", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620991", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "172_skin.fastq.gz", "fastq", 1922415800.0, 38448316.0, "GSM1620991 r1", "0:50", "A:517025440;C:447202733;G:438389966;T:519578929;N:218732", 50, null, null, null, 517025440, 447202733, 438389966, 519578929, 218732, "SRX893445", "SRS859522", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92509, null, 0.08635, null, 0.71747, null, 0.48682, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38396, "SRR1821843", "SRX893444", "SRS859523", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin171 36m 3.75nM roten1 3w rep4", "GSM1620990", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "skin171 36m 3.75nM roten1 3w rep4", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "GSM1620990", "GSM1620990: skin171 36m 3.75nM roten1 3w rep4; Danio rerio; RNA Seq", "GSM1620990", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620990", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "171_skin.fastq.gz", "fastq", 1795802450.0, 35916049.0, "GSM1620990 r1", "0:50", "A:482833890;C:415521773;G:409072097;T:485286706;N:3087984", 50, null, null, null, 482833890, 415521773, 409072097, 485286706, 3087984, "SRX893444", "SRS859523", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91548, null, 0.08856, null, 0.71153, null, 0.48407, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38397, "SRR1821842", "SRX893443", "SRS859524", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin170 36m 3.75nM roten1 3w rep3", "GSM1620989", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "skin170 36m 3.75nM roten1 3w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "GSM1620989", "GSM1620989: skin170 36m 3.75nM roten1 3w rep3; Danio rerio; RNA Seq", "GSM1620989", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620989", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "170_skin.fastq.gz", "fastq", 1844753700.0, 36895074.0, "GSM1620989 r1", "0:50", "A:496864420;C:428934827;G:420432768;T:498397505;N:124180", 50, null, null, null, 496864420, 428934827, 420432768, 498397505, 124180, "SRX893443", "SRS859524", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92475, null, 0.08478, null, 0.71123, null, 0.5082, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38398, "SRR1821841", "SRX893442", "SRS859525", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin169 36m 3.75nM roten1 3w rep2", "GSM1620988", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "skin169 36m 3.75nM roten1 3w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "GSM1620988", "GSM1620988: skin169 36m 3.75nM roten1 3w rep2; Danio rerio; RNA Seq", "GSM1620988", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620988", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "169_skin.fastq.gz", "fastq", 1832979850.0, 36659597.0, "GSM1620988 r1", "0:50", "A:492012429;C:429700408;G:417026859;T:494015583;N:224571", 50, null, null, null, 492012429, 429700408, 417026859, 494015583, 224571, "SRX893442", "SRS859525", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93738, null, 0.07334, null, 0.71936, null, 0.5123, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38399, "SRR1821840", "SRX893441", "SRS859526", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin168 36m 3.75nM roten1 3w rep1", "GSM1620987", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "skin168 36m 3.75nM roten1 3w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:3.75nM roten1|duration:3 weeks", "GSM1620987", "GSM1620987: skin168 36m 3.75nM roten1 3w rep1; Danio rerio; RNA Seq", "GSM1620987", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620987", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "168_skin.fastq.gz", "fastq", 1811602600.0, 36232052.0, "GSM1620987 r1", "0:50", "A:482632140;C:422445561;G:412836586;T:485392445;N:8295868", 50, null, null, null, 482632140, 422445561, 412836586, 485392445, 8295868, "SRX893441", "SRS859526", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.9185, null, 0.07569, null, 0.71368, null, 0.49409, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38400, "SRR1821839", "SRX893440", "SRS859527", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin188 36m control 8w rep4", "GSM1620986", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "skin188 36m control 8w rep4", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "GSM1620986", "GSM1620986: skin188 36m control 8w rep4; Danio rerio; RNA Seq", "GSM1620986", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620986", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "188_skin.fastq.gz", "fastq", 1797310350.0, 35946207.0, "GSM1620986 r1", "0:50", "A:486641633;C:413978398;G:404993329;T:489015902;N:2681088", 50, null, null, null, 486641633, 413978398, 404993329, 489015902, 2681088, "SRX893440", "SRS859527", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91594, null, 0.09158, null, 0.72082, null, 0.51585, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38401, "SRR1821838", "SRX893439", "SRS859528", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin187 36m control 8w rep3", "GSM1620985", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "skin187 36m control 8w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "GSM1620985", "GSM1620985: skin187 36m control 8w rep3; Danio rerio; RNA Seq", "GSM1620985", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620985", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "187_skin.fastq.gz", "fastq", 1951002950.0, 39020059.0, "GSM1620985 r1", "0:50", "A:520701869;C:457907686;G:446812256;T:524196019;N:1385120", 50, null, null, null, 520701869, 457907686, 446812256, 524196019, 1385120, "SRX893439", "SRS859528", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.9211, null, 0.07622, null, 0.71145, null, 0.46944, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38402, "SRR1821837", "SRX893438", "SRS859529", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin186 36m control 8w rep2", "GSM1620984", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "skin186 36m control 8w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "GSM1620984", "GSM1620984: skin186 36m control 8w rep2; Danio rerio; RNA Seq", "GSM1620984", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620984", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "186_skin.fastq.gz", "fastq", 2016117600.0, 40322352.0, "GSM1620984 r1", "0:50", "A:545084574;C:465634059;G:454024180;T:546321124;N:5053663", 50, null, null, null, 545084574, 465634059, 454024180, 546321124, 5053663, "SRX893438", "SRS859529", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91606, null, 0.09659, null, 0.71246, null, 0.52973, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38403, "SRR1821836", "SRX893437", "SRS859530", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin185 36m control 8w rep1", "GSM1620983", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "skin185 36m control 8w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:8 weeks", "GSM1620983", "GSM1620983: skin185 36m control 8w rep1; Danio rerio; RNA Seq", "GSM1620983", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620983", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "185_skin.fastq.gz", "fastq", 1862747450.0, 37254949.0, "GSM1620983 r1", "0:50", "A:506321471;C:429322966;G:419158880;T:507760233;N:183900", 50, null, null, null, 506321471, 429322966, 419158880, 507760233, 183900, "SRX893437", "SRS859530", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.9187, null, 0.09926, null, 0.70755, null, 0.51593, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38404, "SRR1821835", "SRX893436", "SRS859531", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin162 36m control 3w rep3", "GSM1620982", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "skin162 36m control 3w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "GSM1620982", "GSM1620982: skin162 36m control 3w rep3; Danio rerio; RNA Seq", "GSM1620982", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620982", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "162_skin.fastq.gz", "fastq", 2058728600.0, 41174572.0, "GSM1620982 r1", "0:50", "A:545731040;C:487775330;G:479420254;T:545686109;N:115867", 50, null, null, null, 545731040, 487775330, 479420254, 545686109, 115867, "SRX893436", "SRS859531", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93508, null, 0.07365, null, 0.70688, null, 0.4827, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38405, "SRR1821834", "SRX893435", "SRS859532", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin161 36m control 3w rep2", "GSM1620981", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "skin161 36m control 3w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "GSM1620981", "GSM1620981: skin161 36m control 3w rep2; Danio rerio; RNA Seq", "GSM1620981", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620981", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "161_skin.fastq.gz", "fastq", 1897142450.0, 37942849.0, "GSM1620981 r1", "0:50", "A:508043218;C:440251400;G:428510637;T:511046395;N:9290800", 50, null, null, null, 508043218, 440251400, 428510637, 511046395, 9290800, "SRX893435", "SRS859532", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.90644, null, 0.0826, null, 0.71358, null, 0.49573, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38406, "SRR1821833", "SRX893434", "SRS859533", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin160 36m control 3w rep1", "GSM1620980", null, "source name:total RNA extracted from skin|tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "skin160 36m control 3w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:36 month|treatment:n1|duration:3 weeks", "GSM1620980", "GSM1620980: skin160 36m control 3w rep1; Danio rerio; RNA Seq", "GSM1620980", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620980", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "160_skin.fastq.gz", "fastq", 1829692650.0, 36593853.0, "GSM1620980 r1", "0:50", "A:493906226;C:425214476;G:414775189;T:495470406;N:326353", 50, null, null, null, 493906226, 425214476, 414775189, 495470406, 326353, "SRX893434", "SRS859533", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92036, null, 0.09412, null, 0.71484, null, 0.48928, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38407, "SRR1821832", "SRX893433", "SRS859534", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin184 12m control 8w rep3", "GSM1620979", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "skin184 12m control 8w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "GSM1620979", "GSM1620979: skin184 12m control 8w rep3; Danio rerio; RNA Seq", "GSM1620979", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620979", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "184_skin.fastq.gz", "fastq", 1888865600.0, 37777312.0, "GSM1620979 r1", "0:50", "A:510384459;C:436312051;G:428261723;T:512241273;N:1666094", 50, null, null, null, 510384459, 436312051, 428261723, 512241273, 1666094, "SRX893433", "SRS859534", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91542, null, 0.0997, null, 0.70082, null, 0.49633, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38408, "SRR1821831", "SRX893432", "SRS859535", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin183 12m control 8w rep2", "GSM1620978", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "skin183 12m control 8w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "GSM1620978", "GSM1620978: skin183 12m control 8w rep2; Danio rerio; RNA Seq", "GSM1620978", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620978", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "183_skin.fastq.gz", "fastq", 2011969000.0, 40239380.0, "GSM1620978 r1", "0:50", "A:541080523;C:469282719;G:455255702;T:544440274;N:1909782", 50, null, null, null, 541080523, 469282719, 455255702, 544440274, 1909782, "SRX893432", "SRS859535", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.92404, null, 0.104, null, 0.70619, null, 0.47963, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38409, "SRR1821830", "SRX893431", "SRS859536", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin182 12m control 8w rep1", "GSM1620977", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "skin182 12m control 8w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:8 weeks", "GSM1620977", "GSM1620977: skin182 12m control 8w rep1; Danio rerio; RNA Seq", "GSM1620977", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620977", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "182_skin.fastq.gz", "fastq", 1974072600.0, 39481452.0, "GSM1620977 r1", "0:50", "A:532294515;C:457485293;G:447416004;T:532212696;N:4664092", 50, null, null, null, 532294515, 457485293, 447416004, 532212696, 4664092, "SRX893431", "SRS859536", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91699, null, 0.10118, null, 0.71163, null, 0.49086, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38410, "SRR1821829", "SRX893430", "SRS859537", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin159 12m control 3w rep3", "GSM1620976", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "skin159 12m control 3w rep3", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "GSM1620976", "GSM1620976: skin159 12m control 3w rep3; Danio rerio; RNA Seq", "GSM1620976", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620976", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "159_skin.fastq.gz", "fastq", 1887853800.0, 37757076.0, "GSM1620976 r1", "0:50", "A:510759018;C:434865500;G:426519929;T:512499569;N:3209784", 50, null, null, null, 510759018, 434865500, 426519929, 512499569, 3209784, "SRX893430", "SRS859537", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91654, null, 0.1023, null, 0.70607, null, 0.49388, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38411, "SRR1821828", "SRX893429", "SRS859538", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin158 12m control 3w rep2", "GSM1620975", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "skin158 12m control 3w rep2", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "GSM1620975", "GSM1620975: skin158 12m control 3w rep2; Danio rerio; RNA Seq", "GSM1620975", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620975", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "158_skin.fastq.gz", "fastq", 1818559300.0, 36371186.0, "GSM1620975 r1", "0:50", "A:487547586;C:425039144;G:415674047;T:490148749;N:149774", 50, null, null, null, 487547586, 425039144, 415674047, 490148749, 149774, "SRX893429", "SRS859538", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93033, null, 0.09871, null, 0.66103, null, 0.48246, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [38412, "SRR1821827", "SRX893428", "SRS859539", "SRP055573", "PRJNA276667", "RNA seq of zebrafish brain  liver and skin during perturbation with rotenone at young and old age", "GSE66362", "Transcriptome Analysis", "Zebrafish of two different age groups 12 month and 36 month were treated with low amounts of rotenone mild stress and compared to untreated zebrafish. Two different durations were used 3 and 8 weeks. Illumina sequencing HiSeq2000 was applied to generate 50bp single end reads. Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 68 sample: 3 tissues brain  liver  skin; 2 age groups 12 month and 36 month; controls and rotenone treated samples; 2 6 biological replicates for each group", null, "pubmed:27135165", null, "skin157 12m control 3w rep1", "GSM1620974", null, "source name:total RNA extracted from skin|tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "skin157 12m control 3w rep1", "FASTQ files were extracted using Illumina Casava software v1.8.2.", "total RNA extracted from skin", "Tanks with male zebrafish were supplemented with 3.75 nM rotenone or DMSO control for 3 weeks and  8 weeks  respectively. 50% tank water was changed twice a week and fresh Rotenone and DMSO was added to the tanks.", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "Zebrafish Danio rerio were raised under standard conditions.", "tissue:skin|age:12 month|treatment:n1|duration:3 weeks", "GSM1620974", "GSM1620974: skin157 12m control 3w rep1; Danio rerio; RNA Seq", "GSM1620974", null, "1", "Total RNA was isolated using QIAzol Qiagen  Hilden  Germany based on the phenol/chloroform extraction method. postwards the RNA was quantified photometrically with a NanoDrop 1000 PeqLab  Erlangen  Germany and stored at  80 \u00b0C until use. For library preparation an amount of around 2.5 ug of total RNA per sample was processed using Illumina's TruSeqTM RNA Sample Prep Kit Illumina; San Diego; CA  USA following the manufacturer's instruction.", "GEO Accession:GSM1620974", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP055573", null, null, "157_skin.fastq.gz", "fastq", 1855321100.0, 37106422.0, "GSM1620974 r1", "0:50", "A:495304825;C:434601244;G:425046390;T:498975673;N:1392968", 50, null, null, null, 495304825, 434601244, 425046390, 498975673, 1392968, "SRX893428", "SRS859539", "SRA244883", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.91768, null, 0.09102, null, 0.7012, null, 0.49299, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "unknown", "bulk", "unknown", "unknown", null, "Germany", "2015-02-27", "Adult", "Adult", "Skin", "Surface Structure"], [40001, "SRR2751044", "SRX1362114", "SRS1125416", "SRP065208", "PRJNA299585", "Deep sequencing of mRNA from Danio rerio at five different timepoints in three different tissues brain  liver  skin", "GSE74244", "Transcriptome Analysis", "Comparison of temporal gene expression profiles Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 75 samples in sum; 5 age groups 6  12  24  36  42 month; 3 tissues brain  liver  skin; 5 samples per group", null, "pubmed:29382830", null, "NH FLI skin 111", "GSM1915550", null, "source name:total RNA extracted from skin|strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "NH FLI skin 111", "Illumina Casava v1.8.0 software used for extraction of FASTQ files Reads were mapped using Tophat v2.0.6 to the genome taken the annotation into account:  tophat2  p 24  o OUTPUTDIR  g 1   no coverage search   transcriptome index=transcriptome data/Danio rerio.Zv9.73 Danio rerio.Zv9.73 Counting of reads per gene was done using htseq count: htseq count   quiet   stranded=no   idattr=gene id   mode=union SAMFILE Danio rerio.Zv9.73.gtf > OUTPUTFILE Counts were normalized to RPKM values as described in Mortazavi et al. 2008. Gene were excluded from dataset if at least one sample has a RPKM of 0. Genome build: Ensemble Zv9.73 Supplementary files format and content: Excel file includes raw counts and RPKM values for each sample", "total RNA extracted from skin", null, "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", null, "strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "GSM1915550", "GSM1915550: NH FLI skin 111; Danio rerio; RNA Seq", "GSM1915550", null, "1", "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", "GEO Accession:GSM1915550", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP065208", null, null, "skin_42m_DR111.fq.gz", "fastq", 3159101450.0, 63182029.0, "GSM1915550 r1", "0:50", "A:848692326;C:737950616;G:722461308;T:849228405;N:768795", 50, null, null, null, 848692326, 737950616, 722461308, 849228405, 768795, "SRX1362114", "SRS1125416", "SRA306458", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.9327, null, 0.09435, null, 0.70737, null, 0.50049, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2015-10-21", "Adult", "Adult", "Skin", "Surface Structure"], [40002, "SRR2751043", "SRX1362113", "SRS1125415", "SRP065208", "PRJNA299585", "Deep sequencing of mRNA from Danio rerio at five different timepoints in three different tissues brain  liver  skin", "GSE74244", "Transcriptome Analysis", "Comparison of temporal gene expression profiles Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 75 samples in sum; 5 age groups 6  12  24  36  42 month; 3 tissues brain  liver  skin; 5 samples per group", null, "pubmed:29382830", null, "NH FLI skin 109", "GSM1915549", null, "source name:total RNA extracted from skin|strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "NH FLI skin 109", "Illumina Casava v1.8.0 software used for extraction of FASTQ files Reads were mapped using Tophat v2.0.6 to the genome taken the annotation into account:  tophat2  p 24  o OUTPUTDIR  g 1   no coverage search   transcriptome index=transcriptome data/Danio rerio.Zv9.73 Danio rerio.Zv9.73 Counting of reads per gene was done using htseq count: htseq count   quiet   stranded=no   idattr=gene id   mode=union SAMFILE Danio rerio.Zv9.73.gtf > OUTPUTFILE Counts were normalized to RPKM values as described in Mortazavi et al. 2008. Gene were excluded from dataset if at least one sample has a RPKM of 0. Genome build: Ensemble Zv9.73 Supplementary files format and content: Excel file includes raw counts and RPKM values for each sample", "total RNA extracted from skin", null, "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", null, "strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "GSM1915549", "GSM1915549: NH FLI skin 109; Danio rerio; RNA Seq", "GSM1915549", null, "1", "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", "GEO Accession:GSM1915549", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP065208", null, null, "skin_42m_DR109.fq.gz", "fastq", 2638118700.0, 52762374.0, "GSM1915549 r1", "0:50", "A:699567604;C:623885297;G:610871040;T:703405737;N:389022", 50, null, null, null, 699567604, 623885297, 610871040, 703405737, 389022, "SRX1362113", "SRS1125415", "SRA306458", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93453, null, 0.09144, null, 0.7191, null, 0.47437, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2015-10-21", "Adult", "Adult", "Skin", "Surface Structure"], [40003, "SRR2751042", "SRX1362112", "SRS1125418", "SRP065208", "PRJNA299585", "Deep sequencing of mRNA from Danio rerio at five different timepoints in three different tissues brain  liver  skin", "GSE74244", "Transcriptome Analysis", "Comparison of temporal gene expression profiles Jena Centre for Systems Biology of Ageing   JenAge www.jenage.de Overall design: 75 samples in sum; 5 age groups 6  12  24  36  42 month; 3 tissues brain  liver  skin; 5 samples per group", null, "pubmed:29382830", null, "NH FLI skin 108", "GSM1915548", null, "source name:total RNA extracted from skin|strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "NH FLI skin 108", "Illumina Casava v1.8.0 software used for extraction of FASTQ files Reads were mapped using Tophat v2.0.6 to the genome taken the annotation into account:  tophat2  p 24  o OUTPUTDIR  g 1   no coverage search   transcriptome index=transcriptome data/Danio rerio.Zv9.73 Danio rerio.Zv9.73 Counting of reads per gene was done using htseq count: htseq count   quiet   stranded=no   idattr=gene id   mode=union SAMFILE Danio rerio.Zv9.73.gtf > OUTPUTFILE Counts were normalized to RPKM values as described in Mortazavi et al. 2008. Gene were excluded from dataset if at least one sample has a RPKM of 0. Genome build: Ensemble Zv9.73 Supplementary files format and content: Excel file includes raw counts and RPKM values for each sample", "total RNA extracted from skin", null, "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", null, "strain:AB JxTu Tgwt1a:GFP|age:42 month|age category:old 2|tissue:skin", "GSM1915548", "GSM1915548: NH FLI skin 108; Danio rerio; RNA Seq", "GSM1915548", null, "1", "Total RNA from was extracted as described Baumgart et al.  2012; PMID:22487494 Preparation of libraries was done using Illumina\u2019s TruSeq RNA sample prep kit following the manufacturer\u2019s instruction.", "GEO Accession:GSM1915548", "RNA-Seq", "TRANSCRIPTOMIC", "cDNA", "SINGLE", "ILLUMINA", "Illumina HiSeq 2000", null, "SRP065208", null, null, "skin_42m_DR108.fq.gz", "fastq", 2354900150.0, 47098003.0, "GSM1915548 r1", "0:50", "A:629252405;C:552452722;G:540951538;T:632145411;N:98074", 50, null, null, null, 629252405, 552452722, 540951538, 632145411, 98074, "SRX1362112", "SRS1125418", "SRA306458", "GEO", "Leibniz Institute for Age Research - Fritz Lipmann Institute", 1, 0.93959, null, 0.09384, null, 0.71764, null, 0.49807, null, 50, null, "B", null, "usable mapping rate", "illumina", "hiseq_era", "unknown", "cdna_unspecified", "trueseq", "bulk", "unknown", "unknown", null, "Germany", "2015-10-21", "Adult", "Adult", "Skin", "Surface Structure"]], "truncated": false, "filtered_table_rows_count": 256, "expanded_columns": [], "expandable_columns": [], "columns": ["rowid", "run.accession", "experiment.accession", "sample.accession", "study.accession", "bioproject", "study.title", "study.alias", "study.type", "study.abstract", "study.attributes", "study.PMIDs", "sample.description", "sample.title", "sample.alias", "sample.centername", "sample.attributes", "GEOsample.title", "GEOsample.dataprocessing", "GEOsample.source", "GEOsample.treatmentprotocol", "GEOsample.extractprotocol", "GEOsample.growthprotocol", "GEOsample.characteristics", "GEOsample.accession", "experiment.title", "experiment.alias", "experiment.library_name", "experiment.design_description", "experiment.library_construction_protocol", "experiment.attributes", "experiment.library_strategy", "experiment.library_source", "experiment.library_selection", "experiment.library_layout", "experiment.platform", "experiment.instrument_model", "experiment.spot_descriptor", "experiment.study_ref", "run.title", "run.attributes", "run.filename", "run.semantic_name", "run.total_bases", "run.total_spots", "run.alias", "run.read_lengths", "run.base_counts", "run.r1_length", "run.r2_length", "run.r3_length", "run.r4_length", "run.Acount", "run.Ccount", "run.Gcount", "run.Tcount", "run.Ncount", "run.experiment", "run.pool_member", "submission.accession", "submission.srasource", "submission.bioprojectsource", "seqdetective.n_mates", "seqdetective.mapping_rate.mate1", "seqdetective.mapping_rate.mate2", "seqdetective.nofeature_rate.mate1", "seqdetective.nofeature_rate.mate2", "seqdetective.sparsity.mate1", "seqdetective.sparsity.mate2", "seqdetective.pos_strand_rate.mate1", "seqdetective.pos_strand_rate.mate2", "seqdetective.readlen.mate1", "seqdetective.readlen.mate2", "seqdetective.judgement.mate1", "seqdetective.judgement.mate2", "seqdetective.judgement.reason", "platform_family", "instrument_generation", "read_bias", "selection_class", "prep_kit", "sc_or_bulk", "tech_class", "technology", "tech_variant", "submission.bioprojectsource.country", "earliest_date", "devstage_curation", "devstage_curation_coarse", "tissue_curation", "tissue_curation_coarse"], "primary_keys": [], "units": {}, "query": {"sql": "select rowid, [run.accession], [experiment.accession], [sample.accession], [study.accession], bioproject, [study.title], [study.alias], [study.type], [study.abstract], [study.attributes], [study.PMIDs], [sample.description], [sample.title], [sample.alias], [sample.centername], [sample.attributes], [GEOsample.title], [GEOsample.dataprocessing], [GEOsample.source], [GEOsample.treatmentprotocol], [GEOsample.extractprotocol], [GEOsample.growthprotocol], [GEOsample.characteristics], [GEOsample.accession], [experiment.title], [experiment.alias], [experiment.library_name], [experiment.design_description], [experiment.library_construction_protocol], [experiment.attributes], [experiment.library_strategy], [experiment.library_source], [experiment.library_selection], [experiment.library_layout], [experiment.platform], [experiment.instrument_model], [experiment.spot_descriptor], [experiment.study_ref], [run.title], [run.attributes], [run.filename], [run.semantic_name], [run.total_bases], [run.total_spots], [run.alias], [run.read_lengths], [run.base_counts], [run.r1_length], [run.r2_length], [run.r3_length], [run.r4_length], [run.Acount], [run.Ccount], [run.Gcount], [run.Tcount], [run.Ncount], [run.experiment], [run.pool_member], [submission.accession], [submission.srasource], [submission.bioprojectsource], [seqdetective.n_mates], [seqdetective.mapping_rate.mate1], [seqdetective.mapping_rate.mate2], [seqdetective.nofeature_rate.mate1], [seqdetective.nofeature_rate.mate2], [seqdetective.sparsity.mate1], [seqdetective.sparsity.mate2], [seqdetective.pos_strand_rate.mate1], [seqdetective.pos_strand_rate.mate2], [seqdetective.readlen.mate1], [seqdetective.readlen.mate2], [seqdetective.judgement.mate1], [seqdetective.judgement.mate2], [seqdetective.judgement.reason], platform_family, instrument_generation, read_bias, selection_class, prep_kit, sc_or_bulk, tech_class, technology, tech_variant, [submission.bioprojectsource.country], earliest_date, devstage_curation, devstage_curation_coarse, tissue_curation, tissue_curation_coarse from run_metadata where \"experiment.library_strategy\" = :p0 and \"technology\" = :p1 and \"tissue_curation\" = :p2 order by rowid limit 101", "params": {"p0": "RNA-Seq", "p1": "unknown", "p2": "Skin"}}, "facet_results": {"experiment.library_strategy": {"name": "experiment.library_strategy", "type": "column", "hideable": false, "toggle_url": "/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin", "results": [{"value": "RNA-Seq", "label": "RNA-Seq", "count": 256, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?technology=unknown&tissue_curation=Skin", "selected": true}], "truncated": false}, "experiment.library_source": {"name": "experiment.library_source", "type": "column", "hideable": false, "toggle_url": "/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin", "results": [{"value": "TRANSCRIPTOMIC", "label": "TRANSCRIPTOMIC", "count": 256, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_source=TRANSCRIPTOMIC", "selected": false}], "truncated": false}, "experiment.library_selection": {"name": "experiment.library_selection", "type": "column", "hideable": false, "toggle_url": "/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin", "results": [{"value": "cDNA", "label": "cDNA", "count": 188, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_selection=cDNA", "selected": false}, {"value": "Oligo-dT", "label": "Oligo-dT", "count": 39, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_selection=Oligo-dT", "selected": false}, {"value": "PolyA", "label": "PolyA", "count": 24, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_selection=PolyA", "selected": false}, {"value": "RANDOM", "label": "RANDOM", "count": 5, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_selection=RANDOM", "selected": false}], "truncated": false}, "experiment.library_layout": {"name": "experiment.library_layout", "type": "column", "hideable": false, "toggle_url": "/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin", "results": [{"value": "SINGLE", "label": "SINGLE", "count": 192, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_layout=SINGLE", "selected": false}, {"value": "PAIRED", "label": "PAIRED", "count": 64, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.library_layout=PAIRED", "selected": false}], "truncated": false}, "experiment.platform": {"name": "experiment.platform", "type": "column", "hideable": false, "toggle_url": "/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin", "results": [{"value": "ILLUMINA", "label": "ILLUMINA", "count": 246, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&experiment.platform=ILLUMINA", "selected": false}, {"value": "BGISEQ", "label": "BGISEQ", "count": 5, "toggle_url": 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"toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&devstage_curation_coarse=Larval", "selected": false}, {"value": "Embryo", "label": "Embryo", "count": 12, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&devstage_curation_coarse=Embryo", "selected": false}, {"value": "Juvenile", "label": "Juvenile", "count": 12, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&devstage_curation_coarse=Juvenile", "selected": false}, {"value": "Multi-stage", "label": "Multi-stage", "count": 4, "toggle_url": "http://metadata.rnaquarium.org/metadata/run_metadata.json?experiment.library_strategy=RNA-Seq&technology=unknown&tissue_curation=Skin&devstage_curation_coarse=Multi-stage", "selected": false}, {"value": 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