RNA-seq analysis of untreated human differentiating macrophages (rep1)
Indexed cDNA libraries were prepared from 1 ľg of total RNA following the TruSeq RNA Sample Preparation Kit (Illumina, Inc.) protocol. Briefly, the mRNA Poly-A was selected on oligo(dT) magnetic beads and fragmented by divalent cations at 94ĄC for 5 min. Reverse transcription was performed using SuperScript II reverse transcriptase (Life Technologies, Inc.) and random primers. After second strand cDNA synthesis, fragments were end repaired, A-tailed and ligated with indexed adapters. cDNA libraries were purified and size selected after PCR with AMPure XP beads (Beckman Coulter, Inc, Fullerton, C.A USA). The average size of insert was 132ą17 bp.