Trapping gel electrophoresis of end-labeled DNA: an analytical model for mobility and diffusion.
As shown by Ulanovsky, Drouin and Gilbert (Nature 1990, 343, 190-192), the gel electrophoretic migration of DNA is severely reduced by steric trapping when streptavidin is attached to one end of the polyelectrolyte. We present a model that allows us to calculate both the mobility and the diffusion coefficient, hence the resolution factor of the resulting separation. We compare our results to those of Défontaines and Viovy (Electrophoresis 1993, 14, 8-17) and we show that the averages over the molecular conformations must be done carefully. We also show that trapping increases diffusion substantially and that this makes constant-field trapping electrophoresis incapable of increasing the number of bases read per sequencing run. Finally, we conclude that severe trapping may lead to highly anomalous transport behavior where one cannot define a velocity or a diffusion constant.
Slater
G W
GW
Department of Physics, University of Ottawa, Ontario, Canada.
Desruisseaux
C
C
Villeneuve
C
C
Guo
H L
HL
Drouin
G
G
D001426:Bacterial Proteins / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004058:Diffusion; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D008956:Models, Chemical; D019809:Streptavidin; D013997:Time Factors
D001426:Bacterial Proteins; D004247:DNA; D019809:Streptavidin