GSM8751384: H3K4me3_D7-2; Mus musculus; OTHER
Approximately 1×105 cells from each sample were processed using the Hyperactive Universal CUT&Tag Assay Kit for Illumina (Vazyme) following the manufacturer's protocol. Briefly, cells were harvested, and 1×105 cells from each sample were counted. The cells were then incubated with Concanavalin A beads at room temperature for 10 minutes. After removing the liquid, primary antibody was added in ice-cold Antibody Buffer, and the mixture was incubated overnight at 4°C. The following day, the liquid was discarded, and a secondary antibody (Guinea Pig Anti-Rabbit Antibody) in Dig-Wash Buffer was added, followed by a 1-hour incubation at room temperature. After sufficient washing, CUT&Tag pA/G-Tn5 Transposomes were added and incubated at room temperature for 1 hour. Following a final wash, tagmentation was carried out at 37°C for 1 hour. DNA was then extracted using DNA extraction beads. PCR amplification was performed using i7 and i5 Indexed Primers, followed by library cleanup, quantification, and Illumina sequencing. All components except for primary antibody (summarized in key resources table) are included in CUT&Tag Kit.