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Chromatin Immunoprecipitation Assays

Volume 567 of the series Methods in Molecular Biology pp 45-57

Date:

The Fast Chromatin Immunoprecipitation Method

  • Joel NelsonAffiliated withMolecular and Cellular Biology Program, University of Washington
  • , Oleg DenisenkoAffiliated withUW Medicine at Lake Union, University of Washington
  • , Karol BomsztykAffiliated withMolecular and Cellular Biology Program, University of WashingtonUW Medicine at Lake Union, University of Washington

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Abstract

The chromatin immunoprecipitation assay (ChIP assay) has greatly facilitated the recent, dramatic expansion of our knowledge of the protein–DNA interactions involved in regulating gene expression, DNA repair, and cell division. The power of the assay is that it gives a researcher the ability to not only detect a specific protein–DNA interaction in vivo but also determine the relative density of factors along genes or the entire genome. Though powerful, the traditional assay is time consuming (involving 2 days or more) and laborious. With Fast ChIP, we simplified the assay to greatly reduce the time and labor involved. The improved assay is especially useful for studies which involve many samples, including the probing of multiple chromatin factors simultaneously and/or looking at genomic events over several time points. Using Fast ChIP, 24 sheared chromatin samples can be processed to yield PCR-ready DNA in 5 h.

Key words

Chromatin immunoprecipitation ChIP-chip tissue ChIP transcription DNA repair