Journal
NATURE PROTOCOLS
Volume 3, Issue 6, Pages 1018-1025Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2008.66
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Funding
- NSF [MCB-0343934]
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Chromatin immunoprecipitation (ChIP) is a powerful tool for the characterization of covalent histone modifications and DNA-histone interactions in vivo. The procedure includes DNA-histone cross-linking in chromatin, shearing DNA into smaller fragments, immunoprecipitation with antibodies against the histone modifications of interest, followed by PCR identification of associated DNA sequences. In this protocol, we describe a simplified and optimized version of ChIP assay by reducing the number of experimental steps and isolation solutions and shortening preparation times. We include a nuclear isolation step before chromatin shearing, which provides a good yield of high-quality DNA resulting in at least 15 mu g of DNA from each immunoprecipitated sample (from 0.2 to 0.4 g of starting tissue material) sufficient to test >= 25 genes of interest. This simpler and cost-efficient protocol has been applied for histone-modification studies of various Arabidopsis thaliana tissues and is easy to adapt for other systems as well.
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