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Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae
JoVE Journal
Genética
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JoVE Journal Genética
Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae

Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae

DOI:

11:06 min

December 29, 2017

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Capítulos

  • 00:05Título
  • 00:52In Vivo Crosslinking of Proteins to DNA
  • 02:19Making Yeast Lysates
  • 04:08Immunoprecipitation (IP) of Modified Histones
  • 05:11Washing IPs, Eluting Histone-DNA Complexes and Reversing Protein-DNA Crosslinks
  • 07:24Quantitative PCR (qPCR) to Detect Enriched Genomic Regions
  • 08:46Results: ChIP Reveals Histone Modification Status at ant Genomic Region
  • 10:17Conclusion

Summary

Tadução automática

Here, we describe a protocol for chromatin immunoprecipitation of modified histones from the budding yeast Saccharomyces cerevisiae. Immunoprecipitated DNA is subsequently used for quantitative PCR to interrogate the abundance and localization of histone post-translational modifications throughout the genome.

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