CAPILLARY ELECTROPHORESIS OF DNA RESTRICTION FRAGMENTS WITH SOLUTIONS OF ENTANGLED POLYMERS

Separation of DNA restriction fragments in dilute buffer solutions of network forming polymers such as linear polyacrylamide (PAA), hydroxyethyl cellulose (HEC) and polyvinylalcohol (PVA) in phosphate buffer were investigated. PVA in buffers already became inhomogeneous after a few separations with resultant deterioration of resolution. Hydroxylic polymers, capable of forming suitable networks in buffers, are strongly adsorbed to fused silica surfaces suppressing the electroosmotic flow. These polymers could be applied as surface modifiers in capillaries, filled with buffer media containing other polymers such as PAA. Suppression of the electroosmotic flow by adsorbed HEC at pH 7 was slightly more effective than with PVA but the former coating was less stable due to weaker binding to the fused silica surface. Good separations of restriction fragments could be achieved with solutions of PAA or HEC as separation media after surface pretreatment of fused silica capillaries by either PVA rinsing or coating according to Hjerten [41].