GSM3993219: HNE Z-REX; Danio rerio; RNA-Seq
Euthanized embryos were lysed in Trizol using ceramic hard tissue homogenizing mix (VWR), and RNA was isolated following the manufacturer's protocol.  The quality of the RNA was assessed by Nanodrop spectrophotometry (A260/A280 ratio ~2) and fragment analysis using a Bioanalyzer (Agilent). Library construction and sequencing were performed by the RNA sequencing core (RSC) service at Cornell University. Briefly, from 500 ng total RNA input, polyadenylated RNA was isolated with the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). TruSeq-barcoded RNAseq libraries were generated with the NEBNext Ultra Directional RNA Library Prep Kit (New England Biolabs). Each library was quantified with a Qubit 2.0 (dsDNA HS kit; Thermo Fisher) and the size distribution was determined with a Fragment Analyzer (Advanced Analytical) prior to pooling. Libraries were sequenced on a NextSeq500 instrument (Illumina). At least 20M single-end 81nt reads were generated per library.