GSM9744099: WT_H3K4me3; Mus musculus; OTHER
nuclei isolated from ~500,000 mouse OT-1 CD8+ T cells were immobilized on Concanavalin A-coated magnetic beads (Bangs Laboratories) and permeabilized using wash buffer containing 0.02% digitonin (Promega).  Bead-bound nuclei were incubated with rabbit IgG (Millipore), H3K4me1, H3K4me3, H3K27ac (all from Abcam), H3K4me3 (EpiCypher), and H3K27me3 (Diagenode) antibodies overnight at 4°C. The next day, targeted chromatin digestion was achieved by pAG-MNase (EpiCypher) binding and incubation with CaCl2 at 4°C, followed by CUT&RUN DNA purification using MinElute columns (Qiagen). Libraries from CUT&RUN DNA were prepared using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (New England Biolabs) according to the manufacturer's instructions.