GSM7057331: Jurkat, BHLHE40_LV, stimulated, rep1, H3K27ac; Homo sapiens; ChIP-Seq
25 million cells per sample were pelleted and fixed using 1% formaldehyde and quenched with glycine. Pellets were washed with cold PBS+proteinase, resuspended in lysis buffer, and incubated on ice for 10 minutes. Sample was then diluted up to 1 ml in ChiP dilution buffer, sonicated using Covaris E220, and  incubated overnight with H3K27ac antibody(Active Motif, 39133), GATA3(CST-D13C9), or BHLHE40(Novus Biological NB100-1800) and incubated overnight.  Immunopreciitation was performed using protein G beads, and DNA fragments were purified using AMPure XP beads Eluted fragments were then processed with DNA end-repair(Epicenter Biotech), Klenow a base adding(Klenow from NEB), adaptor ligation, PCR amplification, and purification using AMPure XP beads with final size of ~300 bp.