Method and kit for polynucleotide assay including primer-dependa

Details Method and kit for polynucleotide assay including primer-dependa Method and kit for polynucleotide assay including primer-dependa

1986-05-16

1989-07-25

Warden, Robert J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 91, 435810, 436501, 935 77, 935 78, 536 26, 536 27, 536 28, C12Q 168, C12P 1934, G01N 33566, C07H 1906

Patent

active

048513318

ABSTRACT:
A probe polynucleotide binds to a target nucleotide sequence in the nucleic acid of a biological sample, and then is enzymatically extended in the 3'-direction with a mixture of nucleoside triphosphates including at least one nucleoside triphosphate that has been detectably labeled. After separating extended hybrid from unreacted nucleoside triphosphates, detectably-modified nucleotides which have been incorporated are determined. In some forms, the 3'-terminal nucleotide of the probe polynucleotide is selected to form a matched pair with some sample strands, but a mismatched pair with other sample strands. In such cases, if the primer dependent enzyme used for extension is one lacking 3'-exonuclease activity, then only those hybrids forming such a matched pair will be extended and subsequently determined.

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
patent: 4683202 (1987-07-01), Mullis
Sanger, F. et al., Jour. Mol. Biol., 94:441-448, (1975).
Sanger, F., et al., Proc. Natl. Acad. Sci. USA, 74, No. 12:5463-5467, (1977).
Zimmern, D. et al., Proc. Natl. Acad. Sci., U.S.A., 75 No. 9:4257-4261, (1978).
R. K. Saiki et al., Enzymatic Amplification of B-Globin Genomic Sequences and Restriction Site Analysis for Diagnosis of Sickle Cell Anemia, Science, vol. 230, pp. 1350-1354, (1985).

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