GSM2804482: 32-24ct003; Danio rerio; RNA-Seq
Total RNA was extracted from whole larvae following the QIAzol (Qiagen, Germany) protocol. RNA concentration, purity and quality were measured by NanoDrop 1000 (Thermo Scientific, USA) and TapeStation 2200 (Agilent Technologies, USA). Libraries were built from total RNA using the TruSeq RNA Library Prep Kit v2 (Illumina, USA). After purification with oligo-dT beads, mRNAs were fragmented into an average length of 508-541 base pairs. The first strand of complementary DNA was synthesized with random hexamer primers (Illumina, USA), and the second strand was synthesized by Second Strand Master Mix (Illumina, USA). All 18 libraries were barcoded uniquely, and normalized with the KAPA library quantification kit (Kapa Biosystems, USA). The pooled libraries were denatured following the NextSeq System Denature and Dilute Libraries Guide (Illumina, USA), and loaded at 11 pM on a NextSeq 500 reagen cartridge (Illumina, USA) for 150 cycle, paired-end sequencing.