WGS of HCC1143: S fraction
Cells were cultured in RPMI 1640 supplemented with 15% fetal bovine serum at 37C in a 5% CO2 atmosphere. An asynchronous population of ~50 million cells was fixed in ethanol, treated with RNase A (10mg/mL), and stained with propidium iodide (1mg/mL). Stained cells were flow-sorted on a FACSAria II (BD Biosciences) to isolate 1 million G1 and 1 million S phase cells (Koren et al. 2012). DNA was isolated using the MasterPure DNA Purification Kit (Epicentre MCD85201) and libraries were prepared with the TruSeq DNA PCR-Free Library Prep Kit (Illumina).