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

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 91, 4351723, 435317, 436 63, 436 94, 436501, 436508, 536 27, 536 28, 536 29, 935 17, 935 18, 935 76, 935 77, 935 78, C12Q 168, C12P 1934, C12N 100, C12N 1500, G01N 3348, G01N 3300, G01N 33566, G01N 33564, C07H 2102, C07H 2104

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046831959

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: 4395486 (1983-07-01), Wilson et al.
Gorski et al, "Molecular organization of the HLA-SB Region of the Human Major Histocompatibility Complex and Evidence for Two SB Beta-chain Genes", Proc. Natl. Acad. Sci. USA 81: 3934 (1984).
Boss et al, "Cloning and Sequence Analysis of the Human Major Histocompatibility Complex Gene DC-3beta", Proc. Nat. Acad. Sci. USA 81: 5199 (1984).
Okada et al, "Gene Organization of DC and DX Subregions of the Human Major Histocompatibility Complex", Proc. Natl. Acad. Sci. USA 82: 3410 (1985).
Salser, "Cloning cDNA sequences: A general Technique for Propagating Eukaryotic Gene Sequences in Bacterial Cells", in Genetic Engineering, 1978, Charrabarty (ed.), CRC Press, Inc. Boca Raton, Fla., pp. 53-81.
Gaubatz et al, "Strategies for Constructing Complementary DNA for Cloning", J. Theor. Biol. 95: 679 (1982).
Rossi et al., J. Biol. Chem., 257, 9226-9229 (1982).

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