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Nucleic Acids Research, 1989, Vol. 17, No. 19 7779-7784
© 1989

MOLECULAR BIOLOGY

Mutation detection using nucleotide analogs that alter electrophoretic mobility

J. Stephen Kornher and Kenneth J. Livak*

Central Research & Development Department, E.I. du Pont de Nemours & Co., Inc. Experimental Station, PO Box 80328, Wilmington, DE 19880-0328, USA

*To whom correspondence should be addressed

Received June 14, 1989. Revised August 28, 1989. Accepted August 28, 1989.

A simple primer extension assay has been developed to distinguish homologous DNA segments differing by as little as a single nucleotide. DNA strands are synthesized with one of the four natural nucleotides replaced with an analog that affects electrophoretic mobility. DNAs that are the same length but differ in the number of analog molecules per strand exhibit different mobilities on a sequencing gel. In combination with the polymerase chain reaction (PCR; 1, 2), this method has been used to distinguish mutant and normal alleles of the human insulin receptor gene that differ by a single-base substitution. The method appears to be generally applicable to the detection of any nucleotide polymorphism in any segment of DNA.


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