Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: 5 x 10^6 cells were harvested for transcriptomic analysis via RNA-Seq. Cells were centrifuged and the pellet resuspended in 2%/98% DTT/RLT buffer and stored at -80°C. RNA was extracted with Qiagen's RNeasy mini kit (Qiagen #74104) according to manufacturer's protocol with on-column DNase digestion. RNA libraries for sequencing were prepared using TruSeq Stranded mRNA Sample prep kit with 96 dual indexes (Illumina, CA, USA) according to the manufacturer’s instructions with the following changes. The protocols were automated using an Agilent NGS workstation (Agilent, CA, USA) using purification steps as previously described (Borgström et al., 2011; Lundin et al., 2010). Libraries were clustered using cBot and sequenced on HiSeq2500 (HiSeq Control Software 2.2.38/RTA 1.18.61) with a 2x126 setup in RapidHighOutput mode.