MSTO211H DMSO CUT&RUN H3K27ac replicate 1
CUT&RUN libraries were generated with the CUTANA ChIC/CUT&RUN kit version 4 (EpiCypher, 12-1048) and the CUT&RUN library prep kit (EpiCypher, 14-1001&14-1002) according to their instructions. MSTO211H cells were cultured and treated as described above in 10-cm plates. Cells were harvested by trypsinization and counted with Countess 3 Automated Cell counter (Invitrogen). For one CUT&RUN sample preparation, 0.5 million cells were used to isolate nuclei using the CUTANA Pre-Nuclei Extraction Buffer (EpiCypher, 21-1026a). Nuclei were then bound to beads and incubated with 0.5 g antibodies overnight. The following antibodies were used: IgG (EpiCypher 13-0042), and H3K27ac (EpiCypher 13-0059). Target DNAs bound by antibody were digested with pAG-MNase for 2 hours at 4C and extracted by incubating with stop master mix containing 0.5 ng of Escherichia coli Spike-in DNA at 37C.