title: RNA-seq of mESC: WT
design: The cell pellet was resuspended in 900μl of ice-cold polysome lysis buffer, incubated for 5 minutes at 4°C with continuous rotation (50 rpm), passed through a 23G needle (Braun, 4657640) for minimum 10 times, followed by additional 5 min incubation at 4°C with continuous rotation (50 rpm). The cell lysate was clarified by centrifugation at 3000 g/3min/4°C followed by centrifugation of supernatant from the first step at 10'000g/5min/4°C. 50 µl of clarified lysate from each sample was kept aside for mRNAseq library preparation and was snap frozen and stored at -80 °C. Libraries for RNA-sequencing were prepared using the Trueseq Standard mRNA Illumina kit and sequenced on a NovaSeq 6000 instrument at the genomics facility Basel.