The analysis of DNA restriction fragments and polymerase chain reaction products by dynamic sieving electrophoresis

Abstract

Dynamic sieving electrophoresis (DSE) is a new analytical tool used in capillary electrophoresis for the analysis of nucleic acids. By combining many of the principles of traditional slab gel methods with the use of entangled polymer solutions in surface-modified, thin-walled, fused-silica capillaries, it is possible to perform molecular size determinations of DNA restriction fragments and polymerase chain reaction (PCR) products. Analysis of DNA restriction fragments and PCR products by DSE was performed using a DB-17 surface-modified capillary and an entangled polymer solution of either hydroxyethyl cellulose or hydroxypropyl methylcellulose. DNA restriction fragments and PCR products were monitored at 260 nm. DSE has been shown to have higher resolution and efficiency than traditional slab gel methods. In an automated format, DSE offers speed, resolution, quantitation, and efficiency not available with current slab gel methods.