GSM3854904: p53: OCI-AML3 cells Combination Rep1 ChIP-seq; Homo sapiens; ChIP-Seq
Crosslinking was quenched wih 2.5M Glycine, then chromatin lysates were produced using lysis buffer plus protease inhibitors. Samples were then sonicated on ice for 12 cycles of 30 seconds on, 1 minute off, and debris pelleted.  Input controls were removed from each sample. Sonicated lysates were precleared with magnetic beads, then incubated overnight with antibody-bound beads. ChIP samples were then eluted in 50mM Tris-HCl, pH 8; 10mM EDTA; 1% SDS, and crosslinks were reversed by heating at 65°C for 6-18 hours. TE was added to each ChIP or Input sample. Solutions were incubated with RNaseA for 37 °C for 30 minutes, then with proteinase K at 55 °C for 1-2 hours. DNA samples were then purified using a  Qiagen MinElute PCR purification kit. Libraries were constructed using the NEBNext Ultra DNA Lib Prep kit with NEBNext Multiplex Oligos.