GSM6364289: HBEC3 cells, RKO2,  H3K27me3; Homo sapiens; ChIP-Seq
For RYBP ChIP, cells were fixed with 2.5 mM EGS (ethylene glycol-bis-succinimidyl-succinate) for 45 minutes, then fixed with 1% formaldehyde for 8 minutes; for H2AK119ub1 and H3K27me3, cells were crosslinked with 1% formaldehyde only for 8 minutes, after quenching and cell lysis, chromatin was sonicated using a Covaris E220 Evo sonicator. TruSeq ChIP Sample Preparation Kit (Illumina, IP-202-1012, IP-202-1024) was used according to the manufacturer's instructions to perform ChIP-seq library preparation. 5 ng of DNA was used as starting material for input and IP samples. Libraries were amplified using 14 cycles on the thermocycler. Libraries were validated using the Agilent High Sensitivity DNA Kit.