4.7 Article

Protocol for the fast chromatin immunoprecipitation (ChIP) method

Journal

NATURE PROTOCOLS
Volume 1, Issue 1, Pages 179-185

Publisher

NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2006.27

Keywords

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Funding

  1. NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES [R01DK045978, R37DK045978] Funding Source: NIH RePORTER
  2. NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM045134] Funding Source: NIH RePORTER
  3. NIDDK NIH HHS [DK45978] Funding Source: Medline
  4. NIGMS NIH HHS [GM45134] Funding Source: Medline

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Chromatin and transcriptional processes are among the most intensively studied fields of biology today. The introduction of chromatin immunoprecipitations ( ChIP) represents a major advancement in this area. This powerful method allows researchers to probe specific protein- DNA interactions in vivo and to estimate the density of proteins at specific sites genome- wide. We have introduced several improvements to the traditional ChIP assay, which simplify the procedure, greatly reducing the time and labor required to complete the assay. The simplicity of the method yields highly reproducible results. Our improvements facilitate the probing of multiple proteins in a single experiment, which allows for the simultaneous monitoring of many genomic events. This method is particularly useful in kinetic studies where multiple samples are processed at the same time. Starting with sheared chromatin, PCR- ready DNA can be isolated from 16 - 24 ChIP samples in 4 - 6 h using the fast method.

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