Illumina NovaSeq 6000 paired end sequencing; RNA-seq of wild-type zebrafish embryos developmentally exposed to three addictive drugs
RNA-seq of wild-type zebrafish embryos developmentally exposed to three addictive drugs
Zebrafish embryos were exposed to a drug from 1 dpf to 5 dpf and at the end of the exposure period, larvae were collected as 6 pools of 7 embryos per condition for RNA extraction. Samples were lysed in 110 μl RLT buffer (Qiagen) containing 1.1 μl of 14.3M β-mercaptoethanol (Sigma). The lysate was allowed to bind to 450 μl of Agencourt AMPure XP beads (Beckman Coulter) for 15 minutes. The tubes were left on a magnet (Invitrogen) until the solutions cleared and the supernatant was removed without disturbing the beads. Whilst still on the magnet, beads were washed three times with 70% ethanol and allowed to dry for 20 minutes. Total nucleic acid was eluted from the beads following the manufacturer's instructions and treated with DNase I (NEB, Catalogue number M0303L). Illumina TruSeq Stranded mRNA Sample Prep Kit.