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Analysis of the electrophoretic properties of double‐stranded DNA and RNA in agarose gels at a finite voltage gradient
Author(s) -
Edmondson Stephen P.,
Gray Donald M.
Publication year - 1984
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.360231204
Subject(s) - agarose , electrophoresis , gel electrophoresis of nucleic acids , chemistry , nucleic acid , agarose gel electrophoresis , chromatography , gel electrophoresis , dna , rna , analytical chemistry (journal) , biochemistry , gene
The electrophoretic mobilities of double‐stranded (ds) DNAs and ds RNAs of various lenths, L , were measured in gels of 0.4–1.8% (w/v) agarose at a voltage gradient of 1.0 V/cm. Differences in the electrophoresis of ds DNA and ds RNA are presented and discussed. A general expression is derived that describes the electrophoretic mobility, M , of either type of ds nucleic acid as a function of the gel concentration and the nucleic acid length: M = M ′ 1 ( L / L 0 ) −x − M 2 , where M ′ 1 and L 0 are constants, and x and M 2 depend on the agarose gel concentration. The results obtained by fitting our data with this equation are consistent with the mobilities of nucleic acids in a wide range of gel concentrations, including free electrophoresis in solution and electrophoresis in gles of high agarose concentration in which nuleic acids are expected to reptate through the gel matrix. Finally, various methods of plotting agarose gel electrophoresis data are discussed.