title: GSM3854900: p53: OCI-AML3 cells Nutlin-3a Rep1 ChIP-seq; Homo sapiens; ChIP-Seq
library_construction_protocol: 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.