title: GSM5466407: SK-Mel-147-Control-1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was purified using RNeasy Mini kit (Qiagen) and digested with DNase (Qiagen). Total RNA samples (500ng) were processed with the "QuantSeq 3' mRNA-Seq Library Prep Kit (FWD) for Illumina" (Lexogen, Cat.No. 015) by following manufacturer instructions. RNA Quality scores were 8.7 on average (range 7.4-9.5) when assayed on a PerkinElmer LabChip analyzer. Briefly, library generation is initiated by reverse transcription with oligodT priming, and a second strand synthesis is performed from random primers. Primers from both steps contain Illumina-compatible sequences. Libraries are completed by PCR {This 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).}  cDNA libraries are purified, applied to an Illumina flow cell for cluster generation and sequenced on an Illumina instrument (see below) by following manufacturer's protocols. Illumina NextSeq 550 (with v2.5 reagent kits)