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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.
Affiliation(s): (1) Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
(2) Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
(3) Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
(4) Howard Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
(2) Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
(3) Institut Curie, Centre de Recherche, UMR7147-CNRS, Université, Pierre et Marie Curie, Paris, France
(4) Howard Hughes Medical Institute and Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, USA
Book Title: Meiosis: Volume 1, Molecular and Genetic Methods
Series: Methods in Molecular Biology | Volume: 557 | Pub. Date: Aug-01-2009 | Page Range: 117-142 | DOI: 10.1007/978-1-59745-527-5_9
Subject: Cell Biology
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