GSM6935693: H1hESC_ChIP_GATA3_24h_Rep2; Homo sapiens; ChIP-Seq
Cells were dissociated and fixed in mTeSR1 media with formaldehyde at a final concentration of 1% for 10 minutes. Fixation was quenched with glycine and cells were washed with DPBS. Cells were suspended in high salt lysis buffer and chromatin was sheared using a Bioruptor Plus to sizes of 100-400 bp. Soluble chromatin from approximately 3E+06 cells per ChIP was incubated with primary antibody (Abcam; ab199428) and Protein G Dynabeads at 4 degrees overnight. Dynabeads were then washed and bound chromatin was eluted. Crosslinks were reversed by incubation at 65 degrees overnight, and proteins were digested with Proteinase K. DNA was purified using Zymo ChIP DNA Clean and Concentrator Kit. Libraries were prepared using KAPA Hyper Prep Kit (Kapa Biosystems).