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Influence of λ‐DNA concentration on mobilities and dispersion coefficients during agarose gel electrophoresis
Author(s) -
Pernodet Nadine,
Tinland Bernard
Publication year - 1997
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/(sici)1097-0282(19971005)42:4<471::aid-bip9>3.0.co;2-n
Subject(s) - reptation , chemistry , electrophoresis , dispersion (optics) , electric field , dna , gel electrophoresis , agarose gel electrophoresis , photobleaching , analytical chemistry (journal) , agarose , gel electrophoresis of nucleic acids , fluorescence , chromatography , polymer , optics , physics , quantum mechanics , organic chemistry , biochemistry
Earlier works demonstrated theoretically and experimentally that during gel electrophoresis the mobility μ and the dispersion coefficient D x [reflecting band broadening; G. W. Slater and J. Noolandi (1985) Physics Review Letters, Vol. 55, pp. 1579–1582; T. A. Duke, A. N. Semenov, and J. L. Viovy (1992) Physics Review Letters, Vol. 69, pp. 3260–3263] depend on the chain length, the electric field, and on the gel concentration. Using a Fluorescence Recovery After Photobleaching setup coupled with an electrophoretic cell, we show that they also depend of the DNA concentrationC. Two regimes are observed. The first is analogous to a “dilute” regime in the gel where μ and D x are DNA concentration independent. In the second regime, μ and D x decrease when C increases. These results are explained by DNA‐DNA interactions. As expected the C * concentration, under which measurements must be carried out to avoid this effect, is found to be the same as the overlap concentration C * determined in solution. Using concentrations of the studied DNA lower than its C * , μ and D x show a field dependence in good agreement with the predictions of the Biased Reptation model with Fluctuations. © 1997 John Wiley & Sons, Inc. Biopoly 42: 471–478, 1997