title: GSM4459356: K562_4BKO-3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was purified using an RNeasy plus kit (Qiagen), and was treated by DNase I (Qiagen). Samples were prepared and sequenced according to EA | Q² Solutions standard TS Stranded mRNA sequencing protocol using HiSeq - 2x50bp-PE sequencing on an Illumina sequencing platform. RNA samples were converted into cDNA libraries using the Illumina TruSeq Stranded mRNA sample preparation kit (Illumina # RS-122-2103), and with dUTP utilized in place of dTTP in the second strand master mix. Forked barcode adaptors were attached to the cDNAs via ligation. The resulting molecules were PCR amplified. Since only the second strand included the dUTP base, stalling the polymerase, the only viable template was the first strand, thereby preserving the strand information. Final libraries are quantified, normalized and pooled for sequencing.