Polymerase chain reaction heteroduplex polymorphism analysis by entangled solution capillary electrophoresis.

J Cheng; T Kasuga; K R Mitchelson; E R Lightly; N D Watson; W J Martin; D Atkinson

doi:10.1016/0021-9673(94)80556-3

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PubMed · 7951979

Polymerase chain reaction heteroduplex polymorphism analysis by entangled solution capillary electrophoresis.

J Cheng·T Kasuga·K R Mitchelson·E R Lightly·N D Watson·W J Martin·D Atkinson

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Abstract

Heteroduplex DNA polymorphism analysis (HPA) makes use of conformational polymorphisms to alter electrophoretic mobility of fragments and can be used to detect non-restrictable loci. We have developed a novel application of entangled solution capillary electrophoresis (ESCE) to separate heteroduplex and homoduplex DNA molecules. The addition of ethidium bromide and glycerol to the free solution sieving buffer resulted in the improved peak resolution and good reproducibility. Reannealed polymerase chain reaction products could be used directly for mutation screening and with fully automated ESCE the entire HPA may be completed in less than 30 min including sample handling. This technology could provide a rapid and highly efficient way for screening rare mutations among large numbers of individuals.

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Base Sequence Buffers DNA, Fungal Electrophoresis Ethidium Molecular Sequence Data Nucleic Acid Heteroduplexes Polymerase Chain Reaction Polymorphism, Genetic Journal Article Research Support, Non-U.S. Gov't

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J Cheng, T Kasuga, K R Mitchelson, E R Lightly, N D Watson, W J Martin, D Atkinson. 1994-08-12. Polymerase chain reaction heteroduplex polymorphism analysis by entangled solution capillary electrophoresis.. https://doi.org/10.1016/0021-9673(94)80556-3

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