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Related Concept Videos

Chromatin Immunoprecipitation- ChIP02:36

Chromatin Immunoprecipitation- ChIP

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Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
12.6K

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A Semiautomated ChIP-Seq Procedure for Large-scale Epigenetic Studies
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Sequential ChIP Protocol for Profiling Bivalent Epigenetic Modifications (ReChIP).

Bénédicte Desvoyes1, Joana Sequeira-Mendes1, Zaida Vergara1

  • 1Centro de Biología Molecular Severo Ochoa, CSIC-UAM, Nicolás Cabrera 1, Cantoblanco, 28049, Madrid, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|October 21, 2017
PubMed
Summary
This summary is machine-generated.

This study introduces a sequential chromatin immunoprecipitation (ChIP) protocol, called Re-ChIP, for analyzing multiple histone modifications on single chromatin fibers. This method enables a deeper understanding of gene regulation at the single-cell level in Arabidopsis seedlings.

Keywords:
AntibodyChIPChromatin modificationChromatin-binding proteinCross-linkingEpigeneticsHistone marksSequential chromatin immunoprecipitation (Re-ChIP)

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Plant Science

Background:

  • Chromatin immunoprecipitation (ChIP) identifies histone modifications in cell populations.
  • Current ChIP methods provide population-level insights into chromatin organization and gene regulation.
  • Understanding gene regulation at the single chromatin fiber level is crucial.

Purpose of the Study:

  • To develop and describe a sequential ChIP (Re-ChIP) protocol.
  • To enable the identification of multiple chromatin modifications on the same chromatin fiber.
  • To provide a method for studying gene regulation at the single chromatin fiber level in plants.

Main Methods:

  • A sequential chromatin immunoprecipitation (Re-ChIP) protocol was established.
  • The protocol utilizes two antibodies to distinguish different chromatin modifications.
  • Emphasis was placed on necessary controls for reproducible results.

Main Results:

  • The Re-ChIP protocol allows for the identification of distinct chromatin modifications within the same chromatin fiber.
  • The described protocol yields meaningful and reproducible results.
  • The method is optimized for young Arabidopsis seedlings but adaptable to other plant materials.

Conclusions:

  • Sequential ChIP (Re-ChIP) is a valuable technique for investigating chromatin modifications at the single-fiber level.
  • This protocol advances the study of gene regulation by providing single-fiber resolution.
  • The Re-ChIP method offers a pathway to understand complex epigenetic mechanisms in plants.