GSM6734237: E0771_BrM_B; Mus musculus; RNA-Seq
For RNA-seq Total RNA was extracted using the RNeasy kit (Qiagen) 1 µg of total RNA samples was used as provided by the user. Sample RNA Integrity Numbers were in the range 7.6 to 9.4 when assayed on an Agilent 2100 Bioanalyzer. PolyA+ fraction was purified and randomly fragmented, 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 Stranded mRNA Sample Preparation Part # 15031047 Rev. D" kit (this kit incorporates dUTP during 2nd strand cDNA synthesis, which implies that only the cDNA strand generated during 1st strand synthesis is eventually sequenced). Adapter-ligated library was completed by PCR with Illumina PE primers (9 cycles). The resulting purified cDNA library was applied to an Illumina flow cell for cluster generation and sequenced on an Illumina instrument  by following manufacturer's protocols.