title: GSM6165405: RNA - Retene 20 µM - Replicate 4; Danio rerio; RNA-Seq
library_construction_protocol: 48 hpf larvae were pooled into groups of 9 fish each, briefly anesthetized on ice, and excess water removed. Tissue was immediately homogenized with RNAzol RT (Molecular Research Center) and 0.5mm zirconium oxide beads in a Bullet Blender tissue homogenizer (Next Advance). Homogenate was stored at -80 C until RNA isolation. Total RNA was isolated using a Direct-zol RNA MiniPrep Kit (Zymo Research) with DNAse treatment. mRNA was purified using oligo(dT)-attached magnetic beads and fragmented. cDNA was synthesized using random hexamer-primed reverse transcription, end-repaired, 3' adenlylated and adapters were ligated to 3' ends. cDNA was PCR amplified.