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Chromatin Immunoprecipitation

  • Julio C. Ayala
  • Jorge A. Benitez
  • Anisia J. SilvaEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1839)

Abstract

Chromatin immunoprecipitation (ChIP) measures the physical association between a protein and DNA in the cell. In combination with next-generation sequencing, the technique enables the identification of DNA targets for the corresponding protein across an entire genome. Here we describe the immunoprecipitation of Vibrio cholerae DNA bound to the histone-like nucleoid structuring protein (H-NS) tagged with the Flag epitope. The quality of the DNA obtained in this protocol is suitable for next-generation sequencing. The procedure described herein can be readily adapted to other bacteria and DNA-binding proteins.

Key words

Transcription DNA-binding proteins Immunoprecipitation Chromatin immunoprecipitation Next-generation sequencing Vibrio cholerae 

Notes

Acknowledgments

This work was supported by Public Service Research Grants and a Ruth L. Kirschstein National Research Service Award from the National Institute of Allergy and Infectious Disease, National Institutes of Health, Bethesda, Maryland, USA.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Julio C. Ayala
    • 1
    • 2
  • Jorge A. Benitez
    • 1
  • Anisia J. Silva
    • 1
    Email author
  1. 1.Department of Microbiology, Biochemistry and ImmunologyMorehouse School of MedicineAtlantaUSA
  2. 2.Department of MicrobiologyUniversity of Alabama at BirminghamBirminghamUSA

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