GSM9893683: IgG_1; Homo sapiens; OTHER
CUT&RUN assay for H4R3me2s was performed in human ovarian cancer cells. Cells were immobilized on ConA magnetic beads, permeabilized with digitonin buffer, and incubated with H4R3me2s-specific primary antibody overnight at 4°C. After washing, protein A-MNase was applied for targeted chromatin cleavage. CaCl₂-induced MNase digestion was performed on ice and terminated with EDTA/EGTA stop buffer. Soluble chromatin fragments were eluted, purified, and subjected to Illumina sequencing library preparation. Purified CUT&RUN DNA fragments were end-repaired, A-tailed, and ligated to Illumina sequencing adapters. Adapter-ligated DNA was PCR-amplified with indexed primers, followed by size purification to enrich fragments between 150–500 bp. Final libraries were quality-checked by Qubit quantification and agarose gel electrophoresis. Validated libraries were sequenced on the Illumina platform with paired-end 150 bp reads.