GSM5702571: WT DMSO 2; Homo sapiens; RNA-Seq
Total RNA was extracted from cell pellets using the Agilent Absolutely RNA Miniprep Kit following the manufacturer's instructions Total RNA samples, 250ng with RQS=10 in a labchip analysis, were processed into cDNA sequencing libraries with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015). Briefly, library generation is initiated by reverse transcription with oligodT priming, followed by a random-primed second strand synthesis. Primers from both steps contain Illumina-compatible sequences. Libraries were completed by PCR, and sequenced on an Illumina instrument (see below) by following manufacturer's protocols.  {This library preparation kit generates directional libraries stranded in the sense orientation (the read1 (the only read in single read format) has the sense orientation (--library-type fr-secondstrand in TopHat, --stranded=yes in HTSeq)}