title: GSM8146236: U2OS- AiD DiV A- AsiSI -ER siMePCE XRCC4; Homo sapiens; ChIP-Seq
library_construction_protocol: ChIP-seq: After siRNA depletion and AsiSI-mediated DSB induction, cells were fixed with 1% formaldehyde for 10 minutes at room temperature and quenched with 20 mM Glycine for 5 min. Post quenching, the cells were lysed and chromatin was sonicated using a Bioruptor (Diagenode) instrument. DRIP-seq: After siRNA depletion and AsiSI-mediated DSB induction, cells were harvested and genomic DNA was isolated. Extracted genomic DNA was digested overnight at 37°C using a restriction enzyme cocktail (BsrGI, EcoRI, HindIII, and XbaI NEB), followed by treatment with mock or RNase H. For each DRIP, 8 µg of digested DNAs were subject to immunoprecipitation with 10 µg of S9.6 antibody and eluted. The eluates were sonicated using the bioruptor (Diagenode) and used for library preparation. Libraries were constructed using the NEBNext Ultra II kit following manufacturer's instructions.