POLYMERASE CHAIN-REACTION HETERODUPLEX POLYMORPHISM ANALYSIS BY ENTANGLED SOLUTION CAPILLARY ELECTROPHORESIS

Heteroduplex DNA polymorphism analysis (HPA) makes use of conformation al polymorphisms to alter electrophoretic mobility of fragments and ca n be used to detect non-restrictable loci. We have developed a novel a pplication of entangled solution capillary electrophoresis (ESCE) to s eparate heteroduplex and homoduplex DNA molecules. The addition of eth idium bromide and glycerol to the free solution sieving buffer resulte d in the improved peak resolution and good reproducibility. Reannealed polymerase chain reaction products could be used directly for mutatio n screening and with fully automated ESCE the entire HPA may be comple ted in less than 30 min including sample handling. This technology cou ld provide a rapid and highly efficient way for screening rare mutatio ns among large numbers of individuals.