GSM8730100: B16F10 shRNA OSGEP#2 replicat1; Mus musculus; RNA-Seq
Total RNAs from three independent infections were extracted using TRIzol (Life Technologies) following the manufacturer's protocol Libraries were prepared from 500 ng of total RNA using the Illumina Truseq RNA Sample Preparation kit V2. PolyA RNAs were purified with polyT-coated magnetic beads, chemically fragmented, and used as templates for cDNA synthesis with random hexamers. The cDNA ends were then end-blunted, adenylated at the 3′OH ends, and ligated to indexed adaptors. The adaptor-ligated library fragments were enriched by PCR according to Illumina's protocol and purified. Libraries were validated on the Bioanalyser DNA 1000 chip and quantified by qPCR using the KAPA library quantification kit. Sequencing was performed on a HiSeq2000 with a paired-end 2 × 100 base protocol