GSM8656056: MDA-MB-231, DMSO, biol rep1, H3K4me2 (ChIP); Homo sapiens; ChIP-Seq
The chromatin was prepared from 1-5 x107 cells.The samples were fixed in 1% v/v formaldehyde (Sigma, F8775) at room temperature for 10 minutes, followed by the addition of 125 mM Glycine (Serva, SE39004) for 5 min. The samples were washed twice in PBS and then resuspended in ChIP SDS lysis buffer (100 mM NaCl, 50 mM Tris-Cl pH 8.1, 5 mM EDTA pH 8, 0.2% NaN3, and 0.5% SDS). Nuclei were pelleted for 6 minutes at 1200 rpm. The samples were resuspended in 3 ml icecold IP buffer (100mM Tris-Cl pH 8.6, 100mM NaCl, 5mM EDTA pH 8.0, 0.2 % NaN3, 5% (v/v) Triton 100X) and sonicated with a Branson Digital Sonifier 250, with a large tip for cell lines (30% amplitude, 15 second on/15 second off). DNA libraries were prepared from 5-10 ng of input and IP DNA through an in-house protocol (PMID: 23429716).