GSM7500596: PANC-1 cells, IKE treated, bioreplicate 2; Homo sapiens; RNA-Seq
Total RNA was extracted using the Trizol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. rRNAs were removed from isolated total RNAs to retain mRNAs and ncRNAs, which were subsequently fragmented into short fragments and reverse transcribed into complementary DNA (cDNA) with random primers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTPs and reaction buffer, after which the cDNA fragments were purified with QiaQuick PCR extraction kit (Qiagen, Venlo, The Netherlands), end repaired, poly(A) added, and ligated to Illumina sequencing adapters. Then UNG (Uracil-N-Glycosylase) was used to digest the second-strand cDNA. The digested products were size selected by agarose gel electrophoresis and PCR amplified.