GSM6153488: H9C2_Input_1-mRNA; Rattus; RIP-Seq
Total RNA was treated with RQ1 DNase (Promega) to remove DNA, and the quality and quantity of the purified RNA were determined by measuring the absorbance at wavelengths 260nm/280nm (A260/A280) using Nanodrop One (Thermo). 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 construction by KAPA Stranded mRNA-Seq Kit for Illumina® Platforms (#KK8544). Polyadenylated mRNAs were purified by VAHTS mRNA capture Beads (N401-01) and then fragmented. Fragmented mRNAs were converted into double-stranded cDNAs. Following end repair and poly-A tailing, the cDNAs were ligated to Diluted Roche Adaptor (KK8726). The ligation product was purified and size fractionated to 300-500bps, amplified, purified, quantified, and sequenced after storage at -80℃.