GSM8062649: ZR75, H3K4me3, shCtrl, input; Homo sapiens; ChIP-Seq
MCF-7/ZR-75-1 cells were fixed with 1% formaldehyde and sheared with E220 focused ultrasonicator (Covaris), respectively. Sheared chromatins were mixed with 20% of lysate from MEF cells processed identically as spike-in for normalization. Mixed chromatin were incubated at 4℃ overnight with antibodies. Protein A beads were then incubated with the chromatin and antibody mixture for 2 hours at 4℃. Beads were washed, reverse-crosslinked, and DNA were purified with Qiaquick PCR purification kit (Qiagen). KAPA HyperPrep Kit (Roche) was used to prepare ChIP-Rx libraries.