By: Joaquim Roca1 

| Abstract |
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The electrophoretic velocity of a duplex DNA ring is mainly determined by its overall shape. Consequently, DNA topoisomers
of opposite supercoiling handedness can have identical gel velocity, and topoisomers highly supercoiled cannot be separated
beyond some point. These problems are overcome by two-dimensional agarose gel electrophoresis, which involves two successive
electrophoresis steps in one gel slab. The first and second electrophoresis steps are conducted in orthogonal directions with
different concentrations of DNA intercalating agents. These compounds alter the overall shape of the DNA and, thereby, change
the relative mobility of individual DNA topoisomers.
Book Title: DNA Topoisomerases: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 582 | Pub. Date: Sep-22-2009 | Page Range: 27-37 | DOI: 10.1007/978-1-60761-340-4_3
Subject: Genetics/Genomics
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