Process for amplifying, detecting, and/or cloning nucleic acid s

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

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435 91, 4351721, 435320, 536 27, 935 17, 935 18, C12N 1500, C12P 1934, C07H 2104

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active

048001597

ABSTRACT:
The present invention is directed to a process for amplifying and detecting any target nucleic acid sequence contained in a nucleic acid or mixture thereof. The process comprises treating separate complementary strands of the nucleic acid with a molar excess of two oligonucleotide primers, extending the primers to form complementary primer extension products which act as templates for synthesizing the desired nucleic acid sequence, and detecting the sequence so amplified. The steps of the reaction may be carried out stepwise or simultaneously and can be repeated as often as desired.
In addition, a specific nucleic acid sequence may be cloned into a vector by using primers to amplify the sequence, which contain restriction sites on their non-complementary ends, and a nucleic acid fragment may be prepared from an existing shorter fragment using the amplification process.

REFERENCES:
patent: 4237224 (1980-12-01), Cohen et al.
patent: 4683195 (1987-07-01), Mullis et al.
patent: 4683202 (1987-07-01), Mullis
Rossi et al., J. Biol. Chem. 257: 9226-9229 (1982).
E. Taparowsky et al., Cell, 34:581-586 (1983).

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