title: GSM6599762: primNKWG_5793_p5_15min_JH0017; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was isolated according to Smart-seq2 was performed as described previously (Picelli et.  al., 2014). In brief, cells were lysed in 0.2% Triton X-100 in the presence of RNase inhibitor 40 U/µl RNase inhibitor (Takara) and stored at -80°C until ready to proceed. RNA was denatured, reverse transcribed and amplified for 22 cycles. After reverse transcription, 1 ng cDNA was tagmented, amplified and barcoded following the Illumina Nextera indexing strategy. Final libraries were pooled to 4 nM for sequencing on HiSeq3000/4000. cDNA and library quality was assessed using TapeStation 4150 instrument and D5000HS or D1000 screening tapes (Agilent), respectively. DNA concentration was determined using Qubit dsDNA HS kit and instrument (Invitrogen). cDNA and Illumina Nextera indexed samples were purified using Ampure XP beads (Beckman Coulter) at 1:1 ratio. Sequencing was performed by the Biomedical Sequencing Facility at CeMM at 50bp single-end configuration.