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Nat Protoc. 2008;3(6):1032-45. doi: 10.1038/nprot.2008.68.

A rapid micro chromatin immunoprecipitation assay (microChIP).

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1
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo 0317, Norway.

Abstract

Interactions of proteins with DNA mediate many critical nuclear functions. Chromatin immunoprecipitation (ChIP) is a robust technique for studying protein-DNA interactions. Current ChIP assays, however, either require large cell numbers, which prevent their application to rare cell samples or small-tissue biopsies, or involve lengthy procedures. We describe here a 1-day micro ChIP (microChIP) protocol suitable for up to eight parallel histone and/or transcription factor immunoprecipitations from a single batch of 1,000 cells. MicroChIP technique is also suitable for monitoring the association of one protein with multiple genomic sites in 100 cells. Alterations in cross-linking and chromatin preparation steps also make microChIP applicable to approximately 1-mm(3) fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes approximately 8 h for 16 ChIPs.

PMID:
18536650
DOI:
10.1038/nprot.2008.68
[Indexed for MEDLINE]
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