title: Ribo-seq of mESC: CRISPR-KO of RPL39L
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. Polysome profiling and sequencing of ribosome-protected mRNA fragments were performed by combining the protocols from (Ingolia et al. 2012) and (Hornstein et al. 2016).