Method of detecting mutations in DNA and RNA

Details Method of detecting mutations in DNA and RNA Method of detecting mutations in DNA and RNA

1984-04-23

1987-04-07

Kepplinger, Esther M.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

935 78, 4351723, C12Q 168, C12N 1500

Patent

active

046561277

ABSTRACT:
A method of detecting a mutation of a specific nucleotide base in a target nucleic acid chain comprises: (a) hybridizing a labelled probe to the target to form a hybrid in which one end of the probe is positioned adjacent the specific base; (b) adding a nucleotide derivative, e.g. a thionucleotide, under conditions to cause it to join to the end of the probe if it is complementary to the specific base; (c) digesting the hybrid using an exonuclease enzyme under conditions such that the nucleotide derivative protects the probe from digestion; and observing the presence or absence of label attached to the target. The method can be used to detect mutations even when these are not present at restriction enzyme cleavage sites, and does not require the preliminary steps of restriction digestion, gel electrophoresis and DNA (or RNA) blotting.

REFERENCES:
patent: 4351901 (1982-09-01), Bahl
patent: 4358535 (1982-11-01), Falkow
patent: 4521509 (1985-06-01), Benkovic
Sagi, J. et al, Biochem. and Biophys. Res. Comm., vol. 95, No. 1, 1980, pp. 156-162.
Huang, L. H. et al, Nucleic Acids Research, vol. 10, No. 5, 1982, p. 1579.

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