title: Hybridization thermodynamics of DNA oligonucleotides during microchip capillary electrophoresis.
abstract: Capillary electrophoresis (CE) is a powerful analytical tool for performing separations and characterizing properties of charged species. For reacting species during a CE separation, local concentrations change leading to nonequilibrium conditions. Interpreting experimental data with such nonequilibrium reactive species is nontrivial due to the large number of variables involved in the system. In this work we develop a COMSOL multiphysics-based numerical model to simulate the electrokinetic mass transport of short interacting ssDNAs in microchip capillary electrophoresis. We probe the importance of the dissociation constant, K(D), and the concentration of DNA on the resulting observed mobility of the dsDNA peak, μ(w), by using a full sweep of parametric simulations. We find that the observed mobility is strongly dependent on the DNA concentration and K(D), as well as ssDNA concentration, and develop a relation with which to understand this dependence. Furthermore, we present experimental microchip capillary electrophoresis measurements of interacting 10 base ssDNA and its complement with changes in buffer ionic strength, DNA concentration, and DNA sequence to vary the system equilibria. We then compare our results to thermodynamically calculated K(D) values.
authors: Department of Mechanical Engineering, University of California , Santa Barbara, California 93106, United States.
authors: Thomas M
authors: TM
authors: Wynne
authors: Christopher
authors: C
authors: McCallum
authors: Jackson Travis
authors: JT
authors: Del Bonis-O'Donnell
authors: Pete
authors: P
authors: Crisalli
authors: Sumita
authors: S
authors: Pennathur
mesh_terms: D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000379:methods*; D008962:Models, Theoretical; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009994:Osmolar Concentration; D013816:Thermodynamics
chemical_list: D004247:DNA