Effect of operating variables on the separation of DNA molecules by capillary polyacrylamide gel electrophoresis.

Abstract

The influence of operating conditions on the separation of double stranded DNA molecules in capillary polyacrylamide gel electrophoresis is presented in continuation of previous work. In this study, an equation was derived that relates the effects of electrophoresis variables such as gel concentration, field strength, molecular weight and temperature to the migration velocity. As a model system, phage phi X174 DNA restriction fragments obtained by digestion with HaeIII have been examined. As an illustration of optimized conditions, DNA molecules up to the 10(3) base pair range were separated at high temperature (50 degrees C) in less than 6 min and with efficiencies as high as 5000 plates m-1 s-1.