Anti-ONECUT2 antibody produced in rabbit (Sigma-Aldrich, catalog#)
Sequenced DNA Library
library_strategy
ChIP-Seq
library_source
GENOMIC
library_selection
ChIP
library_construction_protocol
Lysates were clarified from sonicated nuclei and histone-DNA complexes were isolated with antibody. Concentrated DNA ChIP or Input DNA was ligated to specific annealed linkers followed by direct sequencing using the Illumina HiSeq3000, system at the UCLA Clinical Microarray Core. ChIP-Seq libraries were prepared with the ChIP-IT High Sensitivity kit from Active Motif. Briefly, 80% confluent 22Rv1 were cross-linked with 1% formaldehyde at room temperature for 10 min. Chromatin was sonicated to 100-500 bp in a Bioruptor UCD-200 (Diagenode) and incubated overnight with 4μg of antibody (anti-OC2 HPA057058). After RNase A and Proteinase K treatment, ChIP DNA was purified with the column system provided with the kit.