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Native/Denaturing Two-Dimensional DNA Electrophoresis and Its Application to the Analysis of Recombination Intermediates

  • Jessica P. Lao
  • Shangming Tang
  • Neil Hunter
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1054)

Abstract

Two-dimensional (2D) gel electrophoresis employs distinct electrophoretic conditions to better resolve complex mixtures of molecules. In combination with Southern analysis, 2D agarose gel electrophoresis is routinely employed to detect and analyze DNA intermediates that arise during the replication and repair of chromosomes. By separating intermediates into their component single-strands, native/denaturing 2D gels can reveal structure that is not apparent under native conditions alone. Here, we describe a general method for native/denaturing two-dimensional gel electrophoresis and its application to understanding the DNA strand-composition of recombination intermediates formed during meiosis.

Key words

Two-dimensional agarose gel electrophoresis 2D gel Native/denaturing Neutral/alkaline Component strand analysis Southern hybridization Homologous recombination Double-strand break 

Notes

Acknowledgements

This work was supported by National Institutes of Health National Institute of General Medical Sciences (NIH NIGMS) grant GM074223 to N.H. N.H. is an Early Career Scientist of the Howard Hughes Medical Institute.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Jessica P. Lao
    • 1
  • Shangming Tang
    • 1
  • Neil Hunter
    • 1
  1. 1.Howard Hughes Medical Institute and the Departments of Microbiology & Molecular Genetics, Molecular & Cellular Biology and Cell Biology & Human AnatomyUniversity of California DavisDavisUSA

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