Chromatin immunoprecipitation to study protein-DNA interactions in budding yeast

Ezhkova, E., Tansey, William P. (2006) Chromatin immunoprecipitation to study protein-DNA interactions in budding yeast. Methods in Molecular Biology, 313. pp. 225-44. ISSN 1940-6029 (Electronic)1064-3745 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/16118437

Abstract

The accurate replication and expression of genetic information is ultimately governed by the interaction of regulatory proteins with specific sites on chromosomes. In recent years, our understanding of how these interactions occur in vivo has advanced considerably, in large part owing to the widespread application of chromatin immunoprecipitation (ChIP), a technique that allows quantification of protein-DNA interactions within the context of native chromatin. The ChIP assay involves three main steps: (1) chemical crosslinking of protein-DNA complexes in intact cells; (2) recovery of specific proteins by immunoprecipitation; and (3) detection of co-precipitating DNA sequences, usually by the polymerase chain reaction (PCR). Here, we provide a detailed description of a ChIP procedure that is commonly used to detect protein-DNA interactions in the yeast Saccharomyces cerevisiae, and discuss various methods for quantifying co-precipitating DNAs.

Item Type: Paper
Uncontrolled Keywords: Chromatin genetics isolation & purification metabolism Chromatin Immunoprecipitation methods DNA, Fungal genetics/isolation & purification metabolism Mycology methods Polymerase Chain Reaction Saccharomyces cerevisiae genetics metabolism Saccharomyces cerevisiae Proteins/isolation & purification metabolism
Subjects: Investigative techniques and equipment > cloning > PCR
Investigative techniques and equipment > assays > cloning > PCR

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Chromatin dynamics
CSHL Authors:
Communities: CSHL labs > Tansey lab
Watson School > Publications
Depositing User: CSHL Librarian
Date Deposited: 13 Jan 2012 17:11
Last Modified: 19 Sep 2014 16:16
URI: http://repository.cshl.edu/id/eprint/22564

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