GSM9463128: AAVS1; Homo sapiens; RNA-Seq
Cell were centrifugated for 5 min at 2,300 x g at 4°C, resuspended in 1 mL TRIzol (Ambion, Life Technologies) and samples processed by the IRIC Genomic platform. In the case of PABIR1 knockout versus wild type cells, cell concentration was measured using a Beckman Z2 Coulter counter in triplicate. ERCC RNA loading control Spike-in mix (Ambion Ref. 4456740) was diluted 1:20 in water and 20 µL per mL spiked in Trizol prior to addition of 1 mL over each cell pellet. Total RNA was isolated using RNeasy mini kit (Qiagen) according to the manufacturer's instructions. RNA was quantified using Qubit (Thermo Scientific) and quality was assessed with the 2100 Bioanalyzer (Agilent Technologies). Transcriptome libraries were generated using the KAPA RNA HyperPrep (Roche) using poly-A selection (Thermo Scientific).