Detection of single base mismatches of thymine and cytosine residues by potassium permanganate and hydroxylamine in the presence of tetralkylammonium salts.
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 piperidine 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.
Gogos
J A
JA
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Karayiorgou
M
M
Aburatani
H
H
Kafatos
F C
FC
D001482:Base Composition; D001483:Base Sequence; D003596:Cytosine / Q000737:chemistry*; D004252:DNA Mutational Analysis; D005260:Female; D006801:Humans; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology*; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011196:Potassium Permanganate / Q000494:pharmacology*; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology; D013941:Thymine / Q000737:chemistry*
D006898:Hydroxylamines; D000644:Quaternary Ammonium Compounds; D011196:Potassium Permanganate; D019811:Hydroxylamine; D003596:Cytosine; C027917:tetramethylammonium; D013941:Thymine