GSM8365501: siCtrl, replicate 3; Homo sapiens; RNA-Seq
HW8 and ChiPSC12 hiPSCs of similar passage number (±2 passages) were lysed on ice using RLT buffer (Qiagen) and detached by scraping. RNA was isolated using the RNeasy Mini kit (Qiagen) with on-column DNA digestion (Qiagen). REH cells were transfected with siRNA and RNA was extracted after 48 hours. Cells were washed with PBS and dissolved in RLT buffer (Qiagen). RNA was isolated using the RNeasy Mini kit (Qiagen) with on-column DNA digestion (Qiagen). Barcoded libraries were prepared from 0.5 µg of total RNA using the TruSeq RNA Sample Preparation v2 kit (low-throughput protocol, Illumina) and quantified with the Bioanalyzer system (Agilent). 7.5 pM denatured libraries were used as input for cBot (Illumina) and subjected to deep sequencing using the HiSeq 4000 (Illumina) for 101 cycles, with an additional 7 cycles for index reading.