GSM1979835: HT10801b_Input_X2_(20151015_7); Homo sapiens; ChIP-Seq
The HT1080-1b fibrosarcoma cell  line was grown in DMEM media  using standard protocols.  Cells were fixed at a density equal to 2 million/ml using formaldehyde to a final concentration of 1%. Cross-linking was quenched with 125mM glycine. Chromatin was released from cells by incubating in lysis buffer (1% SDS, 10mM EDTA and 50 mM Tris-HCl (pH 8.1)) on ice for 10 min. The resulting chromatin solution was diluted 10 times with IP dilution buffer (1% Triton X-100, 2mM EDTA, 150 mM NaCl and 20mM Tris-HCl (pH8.1)) and digested with Micrococcal nuclease (MNase, Sigma, Cat # N3755) at 37°C for 45 min. Reactions were stopped by addition of EGTA to a final concentration of 20 mM and EDTA to 10 mM. The MNase digested chromatin was sonicated as described before (PMID: 24737864). An aliquot containing 100th of the entire chromatine solution was de-cross-linked at 65 degrees C overnight. De-cross-linked chromatin was subjected to RNase A and Proeinase K treatment followed by DNA extraction.
GEO Accession
GSM1979835