GSM1973705: B11_rx3_strong_ZT15; Danio rerio; RNA-Seq
Larvae were sampled in liquid nitrogen and stored at -80°C until further processing. RNA was extracted with TRIzol (Life Technologies) following the manufacturer’s instructions with minor modifications: 30 larvae were sampled in a volume of 1.5 ml TRIzol reagent. Samples were kept at least overnight at -80°C. In order to disrupt the cell membranes, the samples were passed four times through a syringe with a diameter of 0.45 mm. The cDNA libraries for Sample 1-12 (untreated) were generated using the directional mRNA-seq sample preparation kit (#15018460, Rev.A, October 2010, Illumina) following the manufacturer’s protocol.  mRNAseq libraries of samples 13-20 (treated) were generated following the TruSeq mRNA kit protocol (Illumina). The size and concentration of the generated libraries were evaluated with a Bioanalyzer 2100 and adjusted to a concentration of 10 pM. Cluster and sequencing reactions were performed with the TruSeq SBS kit v3-HS and the paired-end cluster v3-cBot-HS kit.