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Distinction between supercoiled and linear DNA in transverse agarose pore gradient gel electrophoresis
Author(s) -
Wheeler David,
Lin JiaHwei,
Chrambach Andreas
Publication year - 1992
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150130185
Subject(s) - agarose , dna supercoil , gel electrophoresis of nucleic acids , agarose gel electrophoresis , dna , electrophoresis , gel electrophoresis , puc19 , biophysics , chemistry , microbiology and biotechnology , plasmid , chromatography , biology , biochemistry , dna replication
Four species of linear DNA and the first four members of a linking series, generated by treatment of plasmid DNA (PUC19, 2.7 kb) with mitochondrial topoisomerase, were differentiated by transverse agarose pore gradient gel electrophoresis. The experimental curves of migration distance vs. agarose concentration (Ferguson curves) of supercoiled DNA exhibit a steeper trajectory than those of linear DNA of the same size range. As a consequence, the four supercoiled species exhibit an increase in apparent size (relative to linear DNA standards) with increasing agarose concentration. Both the crossing of the Ferguson curves with those of linear standards as well as the apparent size increase with agarose concentration can serve to detect supercoiled plasmid‐sized DNA in mixtures with linear DNA.