GSM5732677: genomic DNA input; Homo sapiens; ChIP-Seq
Cells were rinsed with PBS, collected using cell scrapers, centrifuged at 500 x g for 5 minutes and resuspended in swelling buffer (50 mM Tris pH 8, 2.5 mM EDTA and 30% (v/v) glycerol). Cells were then incubated for 30 minutes in swelling buffer at 4oC, sonicated using Bioruptor (Diagenode) for 2 cycles (HI power, cycles of 30 sec on / 1 min off). Nuclei were spun down at 1000 x g for 5 mins at 4oC, resuspended in sonication buffer (50 mM Hepes pH 7.9, 140 mM NaCl, 1 mM EDTA, 1% Triton X-100, 0.1% Na-deoxycholate and 1% SDS), and sonicated for 13 cycles using the same settings as the previous step. 7.5 ug of SMCHD1 antibody (Abcam # ab31865) was added to 250 ug of precleared chromatin and incubated for 3 hours at RT. Library was constructed using KAPA HyperPrep Kit (Roche #07962363001).