GSM5620969: shNQO1_2st; Rattus norvegicus; RNA-Seq
Total RNAs were treated with RQ1 DNase (Promega) to remove DNA. The quality and quantity of the purified RNA were determined by measuring the absorbance at 260nm/280nm (A260/A280) using smartspec plus (BioRad). RNA integrity was further verified by 1.5% agarose gel electrophoresis. For each sample, 1 μg of total RNA was used for RNA-seq library preparation with KAPA Stranded mRNA-Seq Kit for Illumina® Platforms (#KK8544). Polyadenylated mRNAs were purified with VAHTS mRNA capture Beads (N401-01), and fragmented mRNAs were converted into double strand cDNA. Following end repair and A tailing, the DNAs were ligated to Diluted Roche Adaptor (KK8726). Ligation products corresponding to 300-500bps were amplified, purified, quantified, and stored at -80 ℃ before sequencing.