Electrophoresis of end-labeled DNA: theory and experiment.
The dynamic behavior of end-labeled DNA during free-solution electrophoresis is investigated using a simple dumbbell model for the labeled DNA. We study the effect of the applied field, label size, and chain stiffness on DNA conformation and electrophoretic mobility. High applied fields are predicted to magnify the size-dependence of mobility and to yield a nonmonotonic dependence of electrophoretic mobility on applied field. The effectiveness of leveraging label size and DNA chain stiffness for improving resolution is also discussed in the context of DNA deformation. To evaluate the most salient model predictions, we use capillary electrophoresis experiments to characterize the size- and field-dependent mobility of dsDNA fragments (300 bp-2 kbp) end-functionalized with streptavidin. Our experimental results are found to be in generally good accord with expectations based on the dumb-bell model. We discuss implications of these findings for fast, size-based separation of DNA in free solution.
Lau
Henry W
HW
School of Chemical and Biomolecular Engineering, Ithaca, New York 14853, USA.
Archer
Lynden A
LA
D003198:Computer Simulation; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects*; D004574:Electromagnetic Fields; D004586:Electrophoresis / Q000379:methods*; D008956:Models, Chemical; D009038:Motion; D019809:Streptavidin / Q000737:chemistry* / Q000528:radiation effects*; D013314:Stress, Mechanical
D004247:DNA; D019809:Streptavidin