GSM3598105: AKI_DMSO_Rep1; Danio rerio; RNA-Seq
70-100 larvae were pooled and treated with DTT and Collagenase I to disintegrate tissue. Then, they were titurated with p100 to reveal pronephros. GFP+pronephros were dissected and collected. Trizol based RNA isolation was used using RNAeasy Micro Kit (QIAGEN) TruSeq Stranded mRNA kit was used according to the manufacturer's protocol. Fragmentation was performed for 8 minutes. 15 PCR amplification provided selective enrichment of DNA with adapters ligated to both ends and was followed by library quantity and quality assessment by fluorometric assay (Qubit) and Agilent DNA 1000 TapeStation assay respectively. Final libraries were normalized to 10 nM, pooled and diluted. Flowcells for the NextSeq 500 were seeded with 1.8 pM denatured library for automated cluster formation and 2 x 75 paired end sequencing, approximately 30-40 million reads per sample. Four samples of the first biological replicates were pooled and sequenced on a single flow cell. Second and third replicates were pooled and ran on 2 flow cells.
GEO Accession
GSM3598105