GSM5240746: 24hr_k27ac_rep2; Homo sapiens; OTHER
Cells were washed and collected in PBS. The cell pellet was resuspended in RT PBS and fixed for 5min with 1% formaldehyde before quenching with 0.125M Glycine. 5000 fixed drosophila sg4 cells (0.05%) were added as a spike-in control. The cell pellet was obtained, washed in cold PBS and incubated in swelling buffer. The subsequent nuclear pellet was resuspended in lysis buffer and sonicated to fragments of ~200bp. One tenth of the sample was taken as input control. Samples were incubated overnight with the required antibody, 5ul of H2AV drosophila Ab and blocked ProteinA dynabeads. IPs were stingically washed. IPs were tagmented (Nextera DNA Library Prep Kit) and tagmented DNA eluted off the beads. The eluted IPs, and Input controls were decrosslinked, RNase and proteinase K treated. The input controls were tagmented. Nextera DNA library prep kit and Nextera index kit (Illumina)