RNA-Seq of Mus Musculus: 8-week male cortex
Total RNA was extracted using Trizol. mRNA was enriched by Oligo(dT) beads. Then the enriched mRNA was fragmented into short fragments and reverse transcripted into cDNA with random primers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTP and buffer. Then the cDNA fragments were purified with QiaQuick PCR extraction kit, end repaired, poly(A) added, and ligated to Illumina sequencing adapters. The ligation products were size selected by agarose gel electrophoresis, PCR amplified, and sequenced using Illumina HiSeq2500