GSM7295628: Y0RSV_termB2019_S1; Homo sapiens; RNA-Seq
WD-PNEC cultures at 24, 48, 72, 96 hpi were trypsinized in 200 μL 0.05% Trypsin-ethylenediaminetetraacetic acid (EDTA) (v/v).  Once cells were singular and detached as determined by light microscopy observation, the cell suspensions was transferred to RNAse free microcentrifuge tubes and the trypsin was inactivated by 400 μL of low glucose DMEM supplemented with 5% FBS (v/v).  The cells were centrifuged at 489 x g and the resulting supernatant carefully removed.  Cells were resuspended in 200 μL ice-cold sterile RNAse-free PBS.  RNA extraction then proceeded according to manufacturer's instructions (High pure RNA isolation kit, Roche diagnostics, Indianapolis, USA).  Purified RNA was stored at -80oC until used. Agilent 2100 Bioanalyzer was used to assess RNA purity and integrity using the RNA 6000 Nano kit (Agilent Technologies, Santa Clara, CA, USA) according to manufacturer's instructions.  Software analysis (Agilent Technologies, version 2.6) calculated RNA concentration and RNA integrity number (RIN) (a measure of RNA integrity) for each sample.  The vast majority of RNA samples demonstrated RIN values between 8.0 and 10. Samples with RIN values <7 were considered of insufficient quality and discarded. KAPA RNA HyperPrep Kit with RiboErase (HRM), using ribosomal depletion to enrich for mRNA