GSM6912147: mESC_HPRT1R_clone2_RNA_rep1; Mus musculus; RNA-Seq
Cells were grown to medium confluency in 6-well plates. Cells were harvested by washing once with PBS, disso-ciating into single-cell suspending with TrypLE Express (ThermoFisher 12604013) and then neutralizing with mESC media. Cells were pelleted at 500 rcf for 5 minutes and RNA was isolated using Qiagen RNeasy Plus Mini kit, resuspending in 350 μL buffer RLT Plus + β-mercaptoethanol, with homogenization using QIAshredder columns (Qiagen 79654). RNA-seq libraries were prepared from 1 μg total RNA using QIAseq FastSelect -rRNA HMR (Qiagen 334386) and QIAseq Stranded RNA kits (Qiagen 180743) according to the manufacturer's protocol.