GSM6510232: WT_Control_R2_RNA-seq; Mus musculus; RNA-Seq
Total RNA was extracted with total RNA (RNeasy Mini Kit, Qiagen) following manufacturer's instructions and treated with DNaseI (Ambion). PolyA+ RNA was purified with the Dynabeads mRNA purification kit (Invitrogen), randomly fragmented and converted to double-stranded cDNA and processed through subsequent enzymatic treatments of end-repair, dA-tailing and ligation to adapters as in Illumina's 'TruSeq RNA Sample Preparation Guide' (Part # 15031047 Rev. D). Adapter-ligated library was completed by limited-cycle PCR with Illumina PE primers and applied to an Illumina flow cell for cluster generation (TruSeq cluster generation kit v5) and sequenced on HiSeq2000 or Illumina HiSeq2500 instrument by following manufacturer's protocols.