title: GSM4490801: H3K4me3_GFP-OE; Mus musculus; ChIP-Seq
library_construction_protocol: Cells were crosslinked by 1% paraformaldehyde for 10 minutes at room temperature and then quenched by 0.125M glycine for 5 minutes. Cells were lysed on ice for 30 minutes with lysis buffer containing 10 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0), 140 mM NaCl, 1% Triton X-100, 0.2% SDS, 0.1% deoxycholic acid. Chromatin DNA was fragmented to around 200-500bp by Diagenode BioruptorPico sonicator for 45 cycles of 30 seconds on and 30 second off, and then incubated overnight with anti-H3K4me3 or H3K27ac or H3K9ac antibody and Dynabead (Life Technologies) at 4C. Next day, immune complexes were washed once with RIPA buffer with 500mM NaCl and once with LiCl wash buffer (10 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0), 250 mM LiCl, 0.5% NP-40, 0.5% deoxycholic acid). DNA was then reverse crosslinked and eluted overnight in elution buffer (10 mM Tris-Cl (pH 8.0), 5 mM EDTA, 300 mM NaCl, 0.5% SDS, 20 mg/ml proteinase K) at 65C. The third day, eluted DNA was purified by AMPure beads (Beckman-Coulter). NEB Next Ultra DNA Library kit was used to prepare library
attributes.tag: GEO Accession
attributes.value: GSM4490801