GSM5606396: PC_H3K27me3_dKO3 [CUT&Tag]; Mus musculus; OTHER
100,000 Cells were FACS sorted into FACS buffer and resuspended in chilled nuclear extraction (NE) buffer (20mM HEPES-KOH at pH7.9, 10mM KCl, 0.5mM spermidine, 0.1% Triton X-100, 20% glycerol) to exact nuclei, which are then bound to Conconavalin A (conA) coated magnetic beads. Following incubations with primary and secondary antibodies, the nuclei-Con A complex was incubated with pA-Tn5 in tagmentaion buffer (20mM HEPES at pH7.5, 300mM NaCl, 0.5mM spermidine, 10mM MgCl2), DNA released and purified, and PCR amplified into a final CUT&Tag library