GSM5866845: U2OS_SLAMseq_G1_I; Homo sapiens; RNA-Seq
Total RNA was isolated from the sorted cells using RNeasy Plus Mini RNA isolation kit (Cat# 74136, Qiagen) and treated with RNase-free DNase (Cat# EN0521, Thermo Fisher Scientific). Finally, the RNA was alkylated with SLAMseq Kinetics kit (Cat# 061.24, Lexogen) as described by the manufacturer. Till this point samples were kept in dark as much as possible. 500ng of RNA was subjected to library preparation using QuantSeq 3' mRNA-Seq Library Prep Kit FWD for Illumina (Cat# 015.24) following manufacturer's Instructions. All the reagents and seath fluid used for flow cytometry were treated with RNase inhibitor RNAsin Ribonuclease Inhibitor (Cat# N2615, Promega).