期刊
NATURE PROTOCOLS
卷 3, 期 6, 页码 1032-1045出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2008.68
关键词
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资金
- Research Council of Norway
- Norwegian Cancer Society
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 similar to 1-mm(3) fresh- or frozen-tissue biopsies. From cell fixation to PCR-ready DNA, the procedure takes similar to 8 h for 16 ChIPs.
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