Native/Denaturing Two-Dimensional DNA Electrophoresis and Its Application to the Analysis of Recombination Intermediates
- 4.1k Downloads
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 wordsTwo-dimensional agarose gel electrophoresis 2D gel Native/denaturing Neutral/alkaline Component strand analysis Southern hybridization Homologous recombination Double-strand break
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.
- 8.Wahls WP, DeWall KM, Davidson MK (2005) Mapping of ssDNA nicks within dsDNA genomes by two-dimensional gel electrophoresis. J Arkansas Acad Sci 59:178Google Scholar
- 18.Green MR, Sambrook J (2012) Molecular cloning: a laboratory manual (fourth edition). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NYGoogle Scholar