GSM1633577: MeCP2 input; Mus musculus; ChIP-Seq
Mouse hypothalamus tissues were fixed in in 1% formaldehyde in PBS for 10 minutes at room temperature. After quenching the fixation with 125 mM glycine in PBS, the samples were homogenized in hypotonic solutions to release the cytoplasmic fractions. Nuclei were collected at 11,000 rpm in a table-top centrifuge. Chromatin was prepared using a method of sonication followed by a brief Micrococcal Nuclease (Worthington) digestion for 5 minutes at 37°C.  Reactions were stopped by addition of EGTA. The chromatin was then collected by spinning at 13,000 rpm in a table-top centrifuge for 10 minutes. All ChIP experiments were performed using anti-GFP antibody.Precipitated chromatin and input samples were then reverse crosslinked and treated with proteinase K. DNA was recovered and eluted in water using a QIAGEN PCR Purification Kit. libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM1633577