title: RNA-seq for mus  musculus: adult male bone marrow derived macrophages RvD1 6h treated
design: cDNA library for  RNA-Seq was generated from 100ng-400ng total RNA using TruSeq RNA Sample  Preparation Kit (Illumina, San Diego, CA, USA) according to the  manufacturers protocol. Briefly poly-A tailed RNA molecules were pulled down  with poly-T oligo attached magnetic beads. Following purification, mRNA was  fragmented with divalent cations at 85C, and then cDNA was generated by  random primers and SuperScript II enzyme (Life Technologies). Second strand  synthesis was performed followed by end repair, single `A` base addition and  ligation of barcode indexed adaptors to the DNA fragments. Adapter specific  PCRs were performed to generate sequencing libraries. Libraries were size  selected with E-Gel EX 2% agarose gels (Life Technologies) and purified by  QIAquick Gel Extraction Kit (Qiagen). Libraries were sequenced on HiSeq 2500  instrument.