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Nucleic Acids Research, 1990, Vol. 18, No. 23 6807-6814
© 1990

Articles

Detection of single base mismatches of thymine and cytosine residues by potassium permanganate and hydroxylamine in the presence of tetralkylammonium salts

Joseph A. Gogos1, Maria Karayiorgou2, Hiroyuki Aburatani2 and Fotis C. Kafatos1,3,*

1Department of Cellular and Developmental Biology, The Biological Laboratories, Harvard University 16 Divinity Avenue, Cambridge, MA 02138 2Center for Cancer Research, Massachusetts Institute of Technology 77 Massachusetts Avenue, Cambridge, MA 02139, USA 3Institute of Molecular Biology and Biotechnology, Research Center of Crete PO Box 1527, Heraklion, 71110, Crete, Greece

* To whom correspondence should be addressed

Received September 26, 1990. Accepted November 7, 1990.

In the presence of tetramethylammonium chloride, potassium permanganate specifically modifies mismatched thymines. Similarly, the modification of mismatched cytosines by hydroxylamine was enhanced by tetraethylammonium chloride. Modification followed by piperldine cleavage permits specific identification of the T and C mismatches and by extension, when the opposite DNA strand is analyzed, of A and G mismatches as well. These reactions can be performed conveniently with DNA immobilized on Hybond M-G paper. We describe conditions that exploit these reactions to detect mismatches, e.g. point mutations or genetic polymorphisms, using either synthetic oligonucleotide probes or PCR amplification of specific genomic DNA sequences.


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