RNA-Seq of Danio rerio: targeted miRNA
Embryonic zebrafish (< 1 hour post fertilization (hpf)) were microinjected with either a miR-203a inhibitor, miR-34, or exposed to 5 M benzo(a)pyrene (BaP). Larval zebrafish, 72 hpf, were collected from 4 replicates in each treatment and larval samples pooled (30 larvae) and flash frozen. Larvae were homogenized and total RNA was extracted using a miRNeasy mini kit (Qiagen). RNA concentrations were measured with a Nanodrop-2000c and RNA integrity assessed with an Agilent 2100 Bioanalyzer. Samples used for downstream library preparation had RNA Integrity Numbers (RIN) > 9.5. RNA was diluted to 1 g and used in the NEBNext Ultra Library Prep Kit for Illumina (New England Biolabs). An Agilent Bioanalyzer 2100 system was used for assessing the quality of libraries. Paired end, 150 bp sequencing was performed on an Illumina NovoSeq platform by Novogene.