Band broadening in gel electrophoresis: scaling laws for the dispersion coefficient measured by FRAP.
We determined quantitatively the band broadening effect during gel electrophoresis by measuring the longitudinal dispersion coefficient Dx, with a fluorescence recovery after photobleaching setup, coupled to an electrophoretic cell. We carried out measurements as a function of the electric field, the average pore size, and the molecular length of DNA fragments. Our results are in good agreement with the predictions of the biased reptation model with fluctuations described by T. A. Duke et al. [(1992) Physics Review Letters, vol. 69, pp. 3260-3263]. This agreement is observed on single-stranded DNA [persistence length approximately equal to 4 nm; B. Tinland et al. (1997) Macromolecules, vol. 30, pp. 5763-5765] in polyacrylamide gels and on double-stranded DNA (persistence length approximately equal to 50 nm) in agarose gels, two systems where the ratio between the average pore size and the Kuhn length is larger than 1.
Tinland
B
B
Institut Charles Sadron, CNRS, Strasbourg, France. tinland@ics.u-strasbg.fr
Pernodet
N
N
Pluen
A
A
D001482:Base Composition; D004247:DNA / Q000032:analysis / Q000737:chemistry*; D053938:DNA Fragmentation; D004277:DNA, Single-Stranded / Q000032:analysis / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008962:Models, Theoretical; D016062:Porosity
D004277:DNA, Single-Stranded; D004247:DNA