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Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells
JoVE 杂志
遗传学
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JoVE 杂志 遗传学
Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells

Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells

DOI:

11:42 min

May 10, 2019

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Chapters

  • 00:04Title
  • 00:38Determination of MNase Digestion Conditions
  • 04:06Reverse Crosslinking, Purification of DNA, and Analysis of Digested Chromatin
  • 06:33Chromatin Immunoprecipitation
  • 09:24Results: Representative Microphotographs, Agarose Gel Analysis, and Occupancy in the AR Genome
  • 10:42Conclusion

Summary

自动翻译

Chromatin immunoprecipitation (ChIP) is a powerful tool for understanding the molecular mechanisms of gene regulation. However, the method involves difficulties in obtaining reproducible chromatin fragmentation by mechanical shearing. Here, we provide an improved protocol for a ChIP assay using enzymatic digestion.

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