GSM9468928: CAL27,FOXK1 sgRNA,rep1; Homo sapiens; OTHER
The HiChIP protocol was performed  as previously described (Mumbach et al., 2016) using  H3K27ac antibody  (Abcam, ab4729) with the following modifications. Samples were sheared using a Covaris E220 using the following parameters: Fill Level = 10, Duty Cycle = 5, PIP = 140, Cycles/Burst = 200, Time = 4 minutes and then clarified by centrifugation for 15 minutes at 16100 rcf at 4° C. We used 7.5 ug of antibody to H3K27ac and captured the chromatin-antibody complex with 60 uL Protein A beads (Thermo Fisher). Qubit quantification following ChIP ranged from 125-150 ng. The amount of Tn5 used and number of PCR cycles performed were based on the post-ChIP Qubit amounts. HiChIP samples were size selected by PAGE purification (200-800 bp) .