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Chromatin immunoprecipitation (ChIP) provides a means of enriching DNA associated with transcription factors, histone modifications, and indeed any other proteins for which suitably characterized antibodies are available. Over the years, sequence detection has progressed from quantitative real-time PCR and Southern blotting to microarrays (ChIP-chip) and now high-throughput sequencing (ChIP-seq). This progression has vastly increased the sequence coverage and data volumes generated. This in turn has enabled informaticians to predict the identity of multi-protein complexes on DNA based on the overrepresentation of sequence motifs in DNA enriched by ChIP with a single antibody against a single protein. In the course of the development of high-throughput sequencing, little has changed in the ChIP methodology until recently. In the last three years, a number of modifications have been made to the ChIP protocol with the goal of enhancing the sensitivity of the method and further reducing the levels of nonspecific background sequences in ChIPped samples. In this chapter, we provide a brief commentary on these methodological changes and describe a detailed ChIP-exo method able to generate narrower peaks and greater peak coverage from ChIPped material.

Original publication

DOI

10.1007/978-1-4939-3724-0_8

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2016

Volume

1443

Pages

119 - 137

Keywords

Androgen receptor, Cancer, Chromatin immunoprecipitation, Exonuclease, Prostate, Chromatin Immunoprecipitation, DNA, Neoplasm, Exonucleases, High-Throughput Nucleotide Sequencing, Humans, Male, Prostatic Neoplasms, Protein Interaction Mapping, Receptors, Androgen, Sequence Analysis, DNA, Tumor Cells, Cultured