GSM4237399: U937 cells, H3ΔNThr22_rep1 (ChIP-seq); Homo sapiens; ChIP-Seq
Cells were resuspended in ChIP lysis buffer (50mM HEPES-KOH pH 7.5, 140 mM NaCl, 1mM EDTA pH8, 1% Triton X-100, 0.1% Sodium Deoxycholate, 0.1% SDS, protease inhibitors) and incubated for 10 mins on ice. Cell lysate was sonicated with Branson Digital Sonifier (Branson) and diluted with RIPA buffer by 10 fold before incubating overnight with antibody/protein-A magnetic complexes. Beads were washed two times with ChIP wash buffer I (20mM Tris pH8, 0.1%SDS, 1% Triton, 0.1% Sodium Deoxycholate, 2mM EDTA, 150mM NaCl) and two times with ChIP wash buffer II ( 20mM Tris pH8, 0.1%SDS, 1% Triton, 0.1% Sodium Deoxycholate, 2mM EDTA, 500 mM NaCl). IPed DNA was eluted from beads, de-crosslinked at 68C and purified using the MinElute PCR Purification Kit (Qiagen). Purified-DNA was subjected for library preparation using the NEBNext Ultra II DNA library prep kit (NEB) following the manufacturer's protocol