GSM5267256: Control ISG-KO A549-ACE2 cells (not infected with SARS-CoV-2); Homo sapiens; OTHER
Sorted and control (non-infected, not IFN-treated) cells were centrifugated (20 min, 2,000g) and resuspended in lysis buffer (NaCI 300 mM, SDS 0.1%, EDTA 10 mM, EGTA 20 mM, Tris 10 mM) supplemented with 1% Proteinase K (Qiagen #19133) and 1% RNAse A/T1 (ThermoFisher #EN0551) and incubated overnight at 65°C. Two consecutive phenol-chloroform (Sigma #P3803-100ML) extractions were performed and DNA was recovered by ethanol precipitation. Nested PCR was performed using the Herculase II Fusion DNA Polymerase (Agilent, #600679). PCR1 products were purified using QIAquick PCR Purification kit (Qiagen #28104). PCR2 products were purified using Agencourt AMPure XP Beads (Beckman Coulter Life Sciences #A63880). DNA concentration was determined using Qubit dsDNA HS Assay Kit (Thermo Fisher #Q32854) and adjusted to 2 nM prior to sequencing.