GSM4453798
GSM4453798: HT1376_H3K27ac-ChIPSeq_2; Homo sapiens; ChIP-Seq
10-20 M cells were cross-linked with 2% FA for 10 mins on ice where Glycine was used to quench the reaction.  Cross-linked chromatin was then extracted with cell lysis buffer (1% SDS, 50 mM Tris-HCl pH 8, 20 mM EDTA) and sonicated to 100-300 bp (Covaris E220).  5 ug chromatin was used as input control and 20ug chromatin was used for pull-down experiments. Fragmented chromatin was incubated with bead-antibody complex with either anti-mouse IgG dynabeads (ThermoFisher, 11201D) or sheep anti-rabbit IgG dynabeads (ThermoFisher, 11203D) with 4ug of anti-H3K27ac (Active motif, 39133) or 8ug of anti-GATA3 (Santa Cruz, sc268),  at 4°C for overnight in rotator. Pulled down chromatin was stringently washed and used for making libraries IP DNA or control inputs were then processed using Kapa Hyper Prep Kit where they were end repaired, adenylated, adaptor ligated, and PCR amplified with KAPA DNA library prepare kit. Pair-end 2x50/2x100 bp high-throughput sequencing was performed using HiSeq/NovaSeq platform (Illumina).
GEO
GSM4453798
GEO Accession
GSM4453798