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Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells
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JoVE Journal Genetica
Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells

Chromatin Immunoprecipitation Assay Using Micrococcal Nucleases in Mammalian Cells

DOI:
10.3791/59375-v

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11:42 min

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May 10, 2019

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,
1Department of Molecular and Cellular Biology,Beckman Research Institute of City of Hope

Capitoli

  • 00:04Titolo
  • 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

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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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Keywords: Chromatin ImmunoprecipitationMicrococcal NucleasesMammalian CellsTranscriptional RegulationMolecular MechanismsCrosslinkingGlycineCell LysisMNase DigestionSonication

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