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

Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae

DOI:
10.3791/57080-v

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11:06 min

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December 29, 2017

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, , ,
1Department of Biological Sciences,University of Maryland

Chapters

  • 00:05Title
  • 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

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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.

Tags

Chromatin ImmunoprecipitationChIPHistone ModificationsSaccharomyces CerevisiaeGene Expression ControlEnvironmental ConditionsChromatin Binding ProteinsEpitope TagsAntibodiesYeast CellsYPD MediumOptical DensityFormaldehydeGlycineChIP Lysis BufferPMSFProtease InhibitorGlass BeadsBead-beating

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