GSM8449676: HCT116, thymidine-mimosine-NoDoxControl, anti-GFP, replicate 1; Homo sapiens; OTHER
Cut&Run was performed as described by Skene et al., 2018 (https://doi.org/10.1038/nprot.2018.015). Each sample used 300,000 cells and included a modified wash buffer and fixation steps adapted from CUTANA Cross-linking Protocol v1.9 from Epicypher. Samples were fixed with 1% fresh paraformaldehyde for 2 min and quenched with 2.5M Glycine at 1:20 dilution before proceeding to wash steps. Overnight incubation at 55C with 2uL proteinase-K (20mg/mL) and 1.6uL 10% SDS per sample was used to reverse cross-links before DNA purification. Libraries were prepared using KAPA Hyper Prep Kit (Ref# 7962347001-KK8502) following the manufacturer's protocol. Adapters used were IDT for Illumina TruSeq UDI indexes (Ref# 20020178). Library amplification was performed using 13-16x cycles. Samples were sequenced on Illumina NovaSeq 6000 platform for PE150 sequencing.