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Targeted Analysis of Chromatin Events (TACE)

  • Jason Sims
  • Changbin Chen
  • Peter Schlögelhofer
  • Marie-Therese KurzbauerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2061)

Abstract

Visualization of meiotic chromatin from pollen mother cells has become an essential technique to study meiosis in the model plant Arabidopsis thaliana. Here we present an advanced cytogenetic method that combines improved immunocytology with chromosome painting, thereby generating a tool to quantitatively analyze localization of proteins to any given genomic region. Proteins involved in different processes such as DNA double-strand break formation and recombinational repair can be visualized on meiotic chromatin with the additional feature of assessing their abundance at specific chromosomal locations.

Key words

Arabidopsis thaliana Chromosome painting FISH Immunocytology Meiosis 

Notes

Acknowledgments

We thank the European Union (FP7-ITN 560 606956), the University of Vienna (I031-B), and the Austrian Science Fund (SFB F3408; P18036) for funding (J.S., P.S., and M.T.K.). C.C. is supported by the National Science Foundation (IOS:1025881 and IOS:1546792) and the Grant-in-Aid program at the University of Minnesota. We furthermore thank Chris Franklin for providing the ASY1 antibody and Sue Armstrong, James Higgins, and Katja Schneider for very helpful technical demonstrations and discussions.

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Copyright information

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

Authors and Affiliations

  • Jason Sims
    • 1
  • Changbin Chen
    • 2
  • Peter Schlögelhofer
    • 1
  • Marie-Therese Kurzbauer
    • 1
    Email author
  1. 1.Max Perutz Labs, Department of Chromosome Biology, Vienna BioCenter (VBC)University of ViennaViennaAustria
  2. 2.Department of Horticultural ScienceUniversity of Minnesota St. PaulSt. PaulUSA

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