GSM7498554: K562, CUX1, rep2; Homo sapiens; ChIP-Seq
Chromatin was fixed from 100x106 gHPRT and gCUX1 transfected K562 cells using 1% formaldehyde for 10 minutes at room temperature and stopped by the addition of 0.125 M glycine. For SMARCA4 ChIP, protein cross linking was performed first.  Cells were washed 3 times with 1X PBS at room temperature. 10 mL of PBS/MgCl2 were added to the cells after final PBS wash. 80 uL of 0.25M DSG-disuccinimidyl glutarate (Thermo Fischer 20593) was added and incubated for 45 minutes at room temperature. Cells were washed 3 times with 1X PBS and followed by DNA crosslinking with 1% formaldehyde as described above. Fixed chromatin was then sonicated (Bioruptor) for 10 minutes in 30 seconds on/off pulses two times for a total of 20 minutes, with vortexing in between. DNA was purified using a PCR purification kit (Qiagen). Libraries were prepped using the Ovation Ultralow Library Kit Tecan Genomics Inc (0344NB-32) and size selected using SPRIselect beads (Beckman Coulter B23317)