GSM6820951: S11 - ETOH WT; Danio rerio; RNA-Seq
Zebrafish were anesthetized in an ice water bath, decapitated, and the bodies were sectioned into four pieces before being transferred to into FastPrep homogenization tubes containing 500 µL TRIzol Reagent (Invitrogen, Carlsbad, California). The contents were sonicated for 3-4 seconds 3 times, before adding additional 250 µL TRIzol. Again, the contents were sonicated for 3-4 seconds 3 times, and 250 µL TRIzol was added (total TRIzol was 1 mL). The contents were then sonicated 3-4 seconds 3-5 times, depending on if the contents were completely sonicated. RNA was isolated using TRIzol Reagent and RNA Clean & Concentrator kit with in-column DNase-I treatment (Zymo Research, Irvine, CA) and stored at -80°C till further processing. Total RNA was used to isolate poly(A) mRNA using NEBNext Poly(A) mRNA Magnetic Isolation Module, and libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, Ipswich, MA). The quantity of library was assayed using Qubit DNA HS assay (Life Technologies, Carlsbad, CA), and quality was analyzed on Bioanalyzer (Agilent Technologies Inc, Santa Clara, CA).