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Meiosis pp 209-234 | Cite as

Stabilization and Electrophoretic Analysis of Meiotic Recombination Intermediates in Saccharomyces cerevisiae

  • Steve D. Oh
  • Lea Jessop
  • Jessica P. Lao
  • Thorsten Allers
  • Michael Lichten
  • Neil Hunter
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 557)

Abstract

Joint Molecule (JM) recombination intermediates result from DNA strand-exchange between homologous chromosomes. Physical monitoring of JM formation in budding yeast has provided a wealth of information about the timing and mechanism of meiotic recombination. These assays are especially informative when applied to the analysis of mutants for which genetic analysis of recombination is impossible, i.e. mutants that die during meiosis. This chapter describes three distinct methods to stabilize JMs against thermally driven dissolution as well as electrophoretic approaches to resolve and detect JMs at two well-characterized recombination hotspots.

Key words

Meiosis homologous recombination double-strand break joint molecule Holliday junction D-loop strand-exchange electrophoresis 

Notes

Acknowledgments

Work in the Hunter lab is supported by NIH NIGMS grant GM074223; work in the Lichten lab is supported by the Intramural Research Program of the National Cancer Institute. The Hunter lab protocols are modified from protocols developed by Tony Schwacha and Nancy Kleckner (4, 10).

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Copyright information

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

Authors and Affiliations

  • Steve D. Oh
    • 1
  • Lea Jessop
    • 2
  • Jessica P. Lao
    • 3
  • Thorsten Allers
    • 2
  • Michael Lichten
    • 2
  • Neil Hunter
    • 1
  1. 1.Departments of Microbiology and Molecular & Cellular BiologyUniversity of CaliforniaDavisUSA
  2. 2.Laboratory of Biochemistry and Molecular Biology, Center for Cancer Research, National Cancer InstituteBethesdaUSA
  3. 3.Departments of Microbiology and Molecular & Cellular BiologyUniversity of CaliforniaDavis

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