GSM5583571: CnT-BS_H3K27me3_250K; Mus musculus; OTHER
CUT&Tag was performed according to previously published protocol (Kaya-Okur et al., 2019) with slight modification, namely using pA-Tn5 transposome assembled with methylated adapter instead. Briefly, harvested cells were washed and then bound to concanavalin A-coated magnetic beads. The cells were permeabilized and incubated with target-specific primary antibody and then with secondary antibody, followed by incubation with pA-Tn5 transposome and tagmentation at target-binding sites. The tagmented DNA was extracted with Qiagen MinElute PCR Purification Kit. In CUT&Tag-BS, the purified CUT&Tag-DNA was subsequently subjected to tagmentation-based bisulfite sequencing library preparation, which includes an oligonucleotide replacement and gap repair step to ensure that each DNA strand is covalently attached to methylated adapters on both ends, followed by bisulfite conversion and PCR amplification to generate sequencing library. While for non-bisulfite-treated CUT&Tag control, the bisulfite conversion step was removed in library preparation.