title: RNA-Seq of U118 cells
design: After total RNA was extracted, eukaryotic mRNA was enriched by Oligo(dT)beads, while prokaryotic mRNA was enriched by removing rRNA by Ribo-ZeroTM MagneticKit (Epicentre, Madison, WI, USA). Then the enriched mRNA was fragmented into shortfragments using fragmentation buffer and reverse transcripted into cDNA with randomprimers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTP andbuffer. Then the cDNA fragments were purified with QiaQuick PCR extraction kit(Qiagen,Venlo, The Netherlands), end repaired, poly(A) added, and ligated to Illumina sequencingadapters.