title: PRO-seq for Pol II occupancy project
design: PRO-seq for Pol II occupancy project
library_construction_protocol: D. melanogaster S2 cells are cultured at 25°C in Schneider's Drosophila medium (Gibco) supplemented with 10% of inactivated fetal bovine serum. For maintenance, cells were split every 3 to 5 days at ~1-2 million cells per mL for no more than 20 passages. Precision run-on sequencing (PRO-seq) was performed as previously described (Judd et al., 2020; Kwak et al., 2013) with slight modifications. For the Drosophila S2 cell samples, 10 million Drosophila S2 cells were used per sample with a spike-in of 0.1 million (1%) DNMT TKO mESCs. For the DNMT TKO mESC samples, 5 million DNMT TKO mESCs were used per sample with a spike-in of 0.05 million (1%) Drosophila S2 cells.