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Plant Meiosis pp 219-236 | Cite as

Chromatin Immunoprecipitation of Meiotically Expressed Proteins from Arabidopsis thaliana Flowers

  • Christophe Lambing
  • Kyuha Choi
  • Alexander R. Blackwell
  • Ian R. HendersonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)

Abstract

During meiosis recombination occurs between homologous chromosomes which can result in reciprocal exchanges of genetic information, called crossovers. Crossover rate is heterogeneous within the genome, with local regions having a significantly higher recombination rate relative to the genome average. These regions are termed hotspots and typically occur with widths of kilobases. Therefore, there is a need to profile recombination factors at a similar resolution during meiosis via techniques such as chromatin immunoprecipitation (ChIP). Here we describe a ChIP protocol, combined with high throughput sequencing (ChIP-seq) optimised for analysis of meiotically expressed proteins in Arabidopsis thaliana flowers. We provide methods to (1) isolate nuclei and prepare the chromatin for shearing, (2) immunoprecipitate DNA molecules cross-linked to a protein of interest, (3) to size-select and purify immunoprecipitated DNA molecules, and (4) to prepare DNA sequencing libraries suitable for high-throughput sequencing. Together, these methods allow the detection of binding sites for meiotic proteins in the Arabidopsis genome at high resolution, which will provide insights into relationships between meiotic chromosome organization, chromatin and recombination.

Key words

Meiosis Chromatin immunoprecipitation Chromosome axis Recombination Arabidopsis 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Christophe Lambing
    • 1
  • Kyuha Choi
    • 1
    • 2
  • Alexander R. Blackwell
    • 1
  • Ian R. Henderson
    • 1
    Email author
  1. 1.Department of Plant SciencesUniversity of CambridgeCambridgeUK
  2. 2.Department of Life SciencesPohang University of Science and TechnologyPohangRepublic of Korea

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