title: GSM9220168: monocytes_CNS_H3K27ac_MG2; Homo sapiens; ChIP-Seq
library_construction_protocol: Chromatin ImmunoPreciptation-seq library preparation was performed as previously described on 1,000,000 cells per cell line per replicate. For H3K27ac ChIP, the cells were fixed in 1% PFA, pelleted at 1000 x g for 5 min at 4°C, and flash frozen. Frozen cell pellets were lysed on ice in ice-cold LB3 (10 mM Tris-HCl pH 7.5, 100 mM NaCl, 1 mM EDTA, 0.5 mM EGTA, 0.1% Na-deoxycholate, 0.5% N-lauroylarcisine, 1X protease inhibitor cocktail). For ChIP-seq, library preparation was performed using the NebNext Ultra II DNA Library Prep Kit (New England Biolabs E7645). Crosslinking was reversed with overnight proteinase K treatment. The DNA was purified using SpeedBeads (Thermo Scientific Fisher 651520505025) in 20% PEG8000, 1.5 M NaCl to final 12% PEG concentration and then amplified using NebNext High-Fidelity 2X PCR Master Mix (New England Biolabs MO541). The final library was purified by gel excision of 200-500 bp fragments and single-end sequenced.