title: BB85_LN CD4_Placebo_D28
design: "Total RNA was extracted from samples using RNAzol RT and was enriched for polyadenylated transcripts by hybridization with oligo-dT(25) magnetic beads.  Enriched polyadenylated RNA was fragmented randomly with heat and divalent cations, and reverse-transcribed from random hexamer primers.  Second strand synthesis was then performed using RNAse-H and DNA Polymerase I.  Double-stranded cDNA was end repaired and A-tailed, and then ligated to a hairpin-loop sequencing adaptor (New England Biolabs).  Libraries were then enriched and barcoded by PCR, validated on a Bioanalyzer, and quantified by qPCR.  Individually libraries were then pooled and quantified again before cluster generation and sequencing."