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Analysis of Meiotic Recombination Intermediates by Two-Dimensional Gel Electrophoresis

  • Jasvinder S. Ahuja
  • G. Valentin BörnerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 745)

Abstract

During meiosis, programmed double strand breaks give rise to crossover and non-crossover recombination products. Meiotic recombination products are formed via several branched intermediates, including single end invasions and double Holliday junctions. Two-dimensional gel electrophoresis provides a sensitive and specific approach for detecting branched recombination intermediates, determining their genetic requirements, and enriching intermediates for further analysis. Here, we describe analysis of branched recombination intermediates in the yeast Saccharomyces cerevisiae by two-dimensional gel electrophoresis. We also provide an introduction to meiotic time-course procedures, stabilization of branched DNA molecules by interstrand crosslinking, extraction of genomic DNA from meiotic cultures, and quantitative analysis of two-dimensional gel blots.

Key words

Joint molecules meiosis recombination two-dimensional gel electrophoresis double Holliday junction single end invasion 

Notes

Acknowledgments

Work in the Börner lab is supported by NIH NIGMS grant GM080715.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of Biological, Geological and Environmental SciencesCenter for Gene Regulation in Health and Disease, Cleveland State UniversityClevelandUSA

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