GSM9039066: DIPG6_H3K4me3; Homo sapiens; OTHER
CUT&RUN was performed using the EpiCypher CUTANA CUT&RUN kit according to the manufacturer's instructions. Briefly, 500,000 viable cells per reaction were washed and permeabilized with 0.01% Digitonin, which was determined to be the minimum concentration for effective permeabilization. Cells were incubated with 0.5 µg antibody, or a 1:50 dilution for SPT16, on a nutator overnight at 4°C. Following washing, pAG-MNase was added to the reactions and CaCl2 was used to activate antibody-bound MNase for targeted chromatin digestion and release of DNA fragments The entire volume of purified DNA (ranging from 1-10 ng) was used as input for NGS library preparation, according to the EpiCypher CUTANA Library Prep Kit instructions.