Nucleic acid sequence amplification method, detection method, an

Details Nucleic acid sequence amplification method, detection method, an Nucleic acid sequence amplification method, detection method, an

1992-04-28

1996-06-11

Horlick, Kenneth R.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 536 2433, C12Q 168, C12P 1934, C07H 2104

Patent

active

055254624

ABSTRACT:
An oligonucleotide is provided, which has at least a base sequence A, which is 10-30 nucleotides in length and homologous to a part of a specific nucleic acid sequence to be amplified, and a base sequence B, which is 10-30 nucleotides in length and complementary to a sequence 3' to the part of the specific nucleic acid sequence to be amplified. Sequences A and B are separated by a spacer region of 0-20 nucleotides. The oligonucleotide is used in an amplification method, wherein the oligonucleotide is mixed with a sample containing a specific nucleic acid sequence to be amplified and allowed to anneal to the specific nucleic acid sequence in the sample. The annealed oligonucleotide acts as a primer in an elongation reaction, whereas any nonannealed oligonucleotide is decomposed, except for at least part of the base sequence A. The elongation product is then denatured to a single strand for use as a template to which the remaining oligonucleotide anneals to form a primer for subsequent elongation. Also provided is a detection method, which allows for detection of a specific nucleic acid sequence in a sample, as well as reagent kits for use in the amplification and detection methods.

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
patent: 4683202 (1987-07-01), Mullis
patent: 4994368 (1991-02-01), Goodman et al.
Kwok et al., Nature 339, 237-238 (1989).
Nelson et al., PNAS 86, 6686-6690 (1989).
Frohman et al., PNAS 85, 8998-9002 (1988).
Wu and Wallace, GENOMICS 4, 560-569 (1989).
Sambrook et al., Molecular Cloning, Cold Spring Harbor Laboratory Press, see pp. 5.73-5.77 (1989).

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