title: GSM8415605: IgG_E; Homo sapiens; ChIP-Seq
library_construction_protocol: Cells were subjected to ChIP-PCR following standard procedures. Briefly, cells were fixed with crosslink solution (HEPES 50mM pH 8.0; NaCl 100mM; EDTA 1mM; EGTA 0.5mM; 0.5% Formaldehyde) for 10 min at room temperature, and incubated with stop solution (10% Glycine in Tris-HCl 10mM pH 8.0). Cells were lysed for 20 min on ice with 10 mM Tris-HCl pH 8.0, 0.25% Triton X-100, 10 mM EDTA, 0.5 mM EGTA, 20 mM β-glycerol-phosphate, 100 mM NA-orthovanadate, 10 mM Na Butyrate and complete protease inhibitor cocktail. The supernatants were sonicated, centrifuged at 13,000 rpm for 15 min, and supernatants were incubated overnight with the FLAG antibody (Merck Life Science) or with anti-mouse IgGs (Millipore). Precipitates were captured with protein A/G-Sepharose, extensively washed, and the crosslink was reverted by incubating the samples with Proteinase K. NEBNext Ultra DNA Library Prep from Illumina