GSM7916306
Cells were then resuspended in buffer A (10mM HEPES PH7.9, 1.5mM MgCl2, 10mM KCl, 0.5% NP40, 0.5mM DTT) and incubated on ice for 10 minutes. Afterward, cells were resuspended in ChIP Lysis buffer (50mM Tris-Cl PH8.0, 5mM EDTA, 0.2% SDS) and sonicated with BioRuptor (Diagenode). Cell lysate was then cleared by centrifugation at 15000 g for 10 minutes at 4_C. Supernatant was taken and mixed 1:1 with ChIP dilution buffer (300mM NaCl, 2% Triton X-100) and then HA-tag antibody (Cell Signaling Technology #3724). Immunocomplex was then pulled-down with ChIP-Grade Magnetic Protein-G beads (CST 9006). Immunoprecipitate was washed extensively and immunocomplex was eluted by ChIP elution buffer (0.1M NaHCO3 and 1%SDS) with shaking at 65_C for 15 minutes. De-crosslinking was then done at 65_C overnight. DNA was purified for Next generation sequencing. A-tailing and index adapters were added. The DNA fragments were purified by Ampure XP Beads and then amplified by PCR. PCR product was purified with Ampure XP Beads again and then analyzed on the Agilent Technologies 2100 bioanalyzer for quality control. Pair-end 150 bases reads were generated on MGISEQ-2000RS(BGI, Shenzhen, China).