Pulsed-field-trapping electrophoresis: a computer simulation study.
Experimental investigations have shown that adding a large, globular and neutral protein (such as streptavidin) at one end of the DNA fragments to be separated by gel electrophoresis strongly affects the dynamics of these molecules, leading to what is known as trapping electrophoresis (TE). In TE, the velocity decreases much more rapidly with DNA molecular size than under normal gel electrophoresis conditions, suggesting that TE may be used to increase the power of separation of polyacrylamide gel electrophoresis. Unfortunately, the bands are broader and fewer readable bands can fit on a single gel slab. Our previous theoretical study of TE also predicted the existence of long-lasting anomalous regimes where one cannot define a velocity or a diffusion constant. These secondary effects of trapping are related to the very broad distribution of detrapping times (the time needed to exit a trap). In order to increase the usefulness of TE, it has been suggested that pulsed fields may help the molecules exit traps more rapidly. In this article, we present a detailed numerical study of pulsed field TE. We conclude that simple pulsed fields alone may not be enough to increase the sequencing power of polyacrylamide TE because the rate of band broadening cannot be controlled. We also report the existence of anomalous regimes in the presence of pulsed fields, a factor that has been previously neglected in analytical models. Other approaches are also proposed.
Desruisseaux
C
C
Department of Physics, University of Ottawa, Ontario, Canada.
Slater
G W
GW
D001426:Bacterial Proteins; D003198:Computer Simulation; D004247:DNA / Q000302:isolation & purification*; D004058:Diffusion; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D008958:Models, Molecular; D019809:Streptavidin; D013997:Time Factors
D001426:Bacterial Proteins; D004247:DNA; D019809:Streptavidin