GSM6016309: ChIP-H3K27ac-Con2; Homo sapiens; ChIP-Seq
ChIP experiment was performed as previously described. Briefly, cell crosslinking was conducted in 1% formaldehyde for 10 minutes and quenched by glycine. Cell nuclear fraction was extracted by LB1 buffer (50 mM Hepes–KOH, pH 7.5; 1 mM EDTA; 140 mM NaCl; 10% Glycerol; Igepal CA-630; 0.25% Triton X-100) followed by washing in LB2 (10 mM Tris–HCL,pH8.0; 200 mM NaCl; 1 mM EDTA; 0.5 mM EGTA) and LB3 buffer (10 mM Tris–HCl, pH 8; 100 mM NaCl; 1 mM EDTA; 0.5 mM EGTA; 0.1% Na–Deoxycholate; 0.5% N-lauroylsarcosine). Chromatin was sheared to 300-500 bp by Diagenode's bioruptor sonicator and then incubated with antibody-conjugated protein A and G beads for overnight at 4°C with rotation. After washing in RIPA buffer (50 mM Tris, pH 7.6, 150 mM NaCl, 1 mM EDTA, 0.1% SDS, 1% Igepal CA630, 0.5% sodium deoxycholate, Protease inhibitor and RNase inhibitor), the beads were eluted in elution buffer (0.1M NaHCO3, 1% SDS and Proteinase K) for 8~16 hours at 65°C. ChIPed DNA was purified by phenol–chloroform extraction and then used for ChIP-qPCR or ChIP-Seq library preparation. ThruPLEX DNA-Seq kit