Illumina HiSeq 2000 sequencing; p53 Chip-Seq in U2OS treated with UV
p53 Chip-Seq in U2OS treated with UV
U2OS cells were maintained in McCoys 5A media  (Invitrogen) 20 J/m2 UVC irradiation treatment for 0, 8 or 16 hr Formaldehyde Cross-linking and chromatin immunoprecipitation assays for this experiment were performed as described by Boyd and Farnham (MCB 29:8389-8299) and Y. Shang et al. (Cell 103:843-852) with some modifications. Formaldehyde concentration used was 1% for 10 minutes. Antibody used for immunoprecipitation was Rabbit p53 (Santa Cruz FL-393) Purified ChIP DNA was used to prepare Illumina multiplexed sequencing libraries. Libraries for Illumina sequencing were prepared following the Illumina-compatible NEXTflex ChIP-Seq Kit (Bioo Scientific).