GSM8342930: CUT&RUN_ZT20_Vehicle_WDR5_2; Mus musculus; OTHER
Tissue or cells were resuspended in a cell lysis buffer and homogenized using a dounce homogenizer. Nuclei were isolated, washed, and counted. 30k nuclei were used per replicate per time point (tissue) or cell line. Nuclei were bound to concavalin-A beads. Nuclei were incubated overnight with primary antibody at 4 degrees celsius, then washed, and incubated with pA/G-MNase for 1 hour at 4 degrees celsius. Residual pA/G-MNase was washed, the sample resupsended in calcium activation buffer and incubated for 30 minutes at 4 degrees celsius. 2X STOP mix containing RNAse A was added to samples, and incubated at 37 degrees celsius for 15 minutes. Supernatant was collected and DNA isolated using column purification prior to library prep. NEB Next Ultra II Kit