Capillary zone electrophoresis of oligonucleotides. Factors affecting separation

Abstract

The influence of various parameters affecting separation of oligonucleotides by capillary zone electrophoresis has been examined. The effects of pH, ionic strength, and various additives, including highly charged cations such as spermine, were studied. Using polycytidines as model compounds, it was demonstrated that pH in the range of 5-8 and ionic strength in the range of 20-200 mmol/l do not influence the separation of oligonucleotides substantially. However, with the addition of spermine to the background electrolyte, migration order was inverted as the effective mobilities of the larger oligonucleotides were greatly decreased. With the addition of spermine and sodium dodecyl sulfate, the separation+ of these model oligonucleotides was also significantly affected. The best separation of a homologous series of polycytidines was obtained with a background electrolyte containing 60 mmol/l histidine, 30 mmol/l glutamic acid, 50 mmol/l sodium dodecyl sulfate and 3 mmol/l spermine.