Gel Electrophoresis Assays for Analyzing DNA Double-Strand Breaks in Saccharomyces cerevisiae at Various Spatial Resolutions (original) (raw)

Abstract

Meiotic recombination is triggered by programmed DNA double-strand breaks (DSBs), which are catalyzed by Spo11 protein in a type II topoisomerase-like manner. Meiotic DSBs can be detected directly using physical assays (gel electrophoresis, Southern blotting, and indirect end-labeling) applied to samples of genomic DNA from sporulating cultures of budding and fission yeast. Such assays are extremely useful for quantifying and characterizing many aspects of the initiation of meiotic recombination, including the timing of DSB formation relative to other events, the distribution of DSBs across the genome, and the influence on DSB formation of mutations in recombination factors and other gene products. By varying the type of gel electrophoresis and other parameters, the spatial resolution of DSB analysis can range from single nucleotides up to whole yeast chromosomes.

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Authors and Affiliations

  1. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
    Hajime Murakami
  2. Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
    Valérie Borde
  3. Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
    Alain Nicolas
  4. Howard Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
    Scott Keeney

Authors

  1. Hajime Murakami
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  2. Valérie Borde
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  3. Alain Nicolas
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  4. Scott Keeney
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Editor information

Editors and Affiliations

  1. Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, York Ave. 1275, New York, 10021-6007, U.S.A.
    Scott Keeney

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Murakami, H., Borde, V., Nicolas, A., Keeney, S. (2009). Gel Electrophoresis Assays for Analyzing DNA Double-Strand Breaks in Saccharomyces cerevisiae at Various Spatial Resolutions. In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 557. Humana Press. https://doi.org/10.1007/978-1-59745-527-5\_9

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