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Nature Protocols 3, 1032–1045 (1 May 2008) | doi:10.1038/nprot.2008.68
A rapid micro chromatin immunoprecipitation assay (ChIP)
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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 (|[mu]|ChIP) protocol suitable for up to eight parallel histone and/or transcription factor immunoprecipitations from a single batch of 1,000 cells. |[mu]|ChIP 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 |[mu]|ChIP applicable to |[sim]|1-mm3 fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes |[sim]|8 h for 16 ChIPs.
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