GSM4437242: uty_ech_1; Mus musculus; ChIP-Seq
Tissues were homogenized and cross-linked in 1% formaldehyde at room temperature for 10 minutes.   The reaction was stopped by adding glycine (125mM), followed by an additional incubation for 5 minutes.   Chromatin was prepared as described, with little modifications (21).  Nuclei were prepared by sequential   incubation and purification by centrifugation on ice for 5 min in buffer A [10 mM Tris-HCl (pH 8), 10mM EDTA  0.25% Triton X-100], and for 30 min in buffer B [10 mM Tris-HCl (pH 8), 1 mM EDTA, 200 mM NaCl] (all buffers  include protease inhibitors). Nuclei were resuspended in sonication buffer [10mMTris-HCl (pH 8), 1mMEDTA,   0.5% SDS, 0.5% Triton X-100, 0.05% NaDOC, 140 mM NaCl] and sonicated to obtain chromatin fragments of an average length of 500 bp. Sonicated chromatin was immunoprecipitated with either rabbit IgG (G2018, Sigma-Aldrich)  or ChIP grade ab4729 anti-H3K27 antibody from Abcam (Cambridge, MA).  The immunoprecipitate was collected using  protein-A/G beads (Santa Cruz Biotechnology). After washes and decrosslinking, DNA was purified using QIAquick columns following manufacturer's directives (Qiagen, Mississauga, On) Immunoprecipitated DNA was used to prepare sequencing libraries using the TruSeq DNA kit   from Illumina, according to the manufacturer's instructions.
GEO Accession
GSM4437242