RNA-Seq of Danio rerio: Pooled larvae
Pooled larval zebrafish (18 pooled larvae for 48 hours post fertilization (hpf) and 12 pooled larvae for 72 and 144 hpf), with 3 replicates for each exposure treatment group, were homogenized in a bead mill with a 5 mm stainless steel ball. Total RNA was extracted using a miRNeasy Mini Kit (Qiagen). A Nanodrop (Thermo Fisher Scientific) was used to measure RNA concentrations and RNA quality and integrity were assessed with an Agilent 2100 Bioanalyzer. Only samples with high-quality RNA Integrity Numbers (RINs) > 9 were used for downstream library preparation. Paired end, 150 bp sequencing was performed on an Illumina NovoSeq 6000 platform by Novogene.