GSM5655527: VCaP_DMSO_2h_2; Homo sapiens; RNA-Seq
Total RNA was isolated from cells using the Direct-zol kit (Zymo, Irvine, CA) and qualuty checked using the Bioanalyzer machine (only samples with RIN>9 was passed for sequencing). RNA-seq libraries were prepared using 200–1,000 ng of total RNA. PolyA+ RNA isolation, cDNA synthesis, end-repair, A-base addition, and ligation of the Illumina indexed adapters were performed according to the TruSeq RNA protocol (Illumina). Libraries were size selected for 250–300 bp cDNA fragments on a 3% Nusieve 3:1 (Lonza) gel, recovered using QIAEX II reagents (QIAGEN), and PCR amplified using Phusion DNA polymerase (New England Biolabs). Library quality was measured on an Agilent 2100 Bioanalyzer for product size and concentration. Paired-end libraries were sequenced with the Illumina HiSeq 2500, (2×100 nucleotide read length) with sequence coverage to 15-20M paired reads.