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Nucleic Acids Research, 1987, Vol. 15, No. 20 8387-8398
© 1987

Articles

Plasmid migration using orthogonal-field-alternation gel electrophoresis

Robin C. Hightower, David W. Metge1 and Daniel V. Santi*

Departments of Biochemistry and Biophysics, and Pharmaceutical Chemistry, University of California San Francisco, CA 94143 1Department of Microbiology, San Francisco State University San Francisco, CA 94132, USA

*To whom correspondence should be addressed

Received June 19, 1987. Revised September 14, 1987. Accepted September 14, 1987.

The migration properties of a series of supercoiled plasmids ranging in size from 4 to 16 kilobases (kb) have been analyzed by orthogonal-field-alternation gel electrophoresis (OFAGE). These circular DNAs enter the gel and are well resolved. Unlike linear DNA molecules, the relative mobilities of these plasmids are constant over a wide range of pulse times, from 10 to 120 seconds, as well as over a broad range of total running times, from 6 to 24 hours. Electrophoresis of supercoiled, relaxed, and nicked open circular forms as well as topoisomers of pBR322 shows that the extent of supercoiling has a dramatic effect on plasmid migration on OFAGE. Several practical applications for exploiting the different migration properties of circular and linear DNA molecules on OFAGE are presented.


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