GSM5551321: 4hr Vehicle H3K4me3; Homo sapiens; ChIP-Seq
Chromatin complexes were eluted in 100 μL of elution buffer (10 mmol/L Tris-HCl [pH 7.6], 1 mmol/L EDTA, 250 mmol/L NaCl, and 0.3% SDS) and incubated at 37°C for 1 hour with shaking. For protein degradation and reverse cross-linking, 5 μL of Proteinase K solution (Life Technologies) was added to the eluates and incubated at 37°C for 1 hour, followed by incubation at 65°C for 6 hours. DNA was purified with the QIAquick PCR Purification Kit (Qiagen, Valencia, Calif), eluted in 30 μL of molecular biology grade water (Sigma, St Louis, Mo), and processed for qPCR before sequencing. Libraries were created, as described previously,38 and sequenced by the CCHMC Gene Discovery and Genetic Variation Core.