Promoter ligation activated transcription amplification of nucle

Details Promoter ligation activated transcription amplification of nucle Promoter ligation activated transcription amplification of nucle

1990-05-16

1993-03-16

Moskowitz, Margaret

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 91, 436501, 436 94, 935 77, 935 78, C12Q 168, C12P 1934, G01N 33566, G01N 3348

Patent

active

051943700

ABSTRACT:
This invention discloses a scheme for producing nucleic acid end products that are functionally or exactly identical to the starting products, thereby resulting in exponential amplification of a desired nucleic acid sequence. Specifically, sequences are cycled between RNA and DNA forms using the following basic steps: (1) a T7 RNA polymerase promoter is ligated onto a single-stranded DNA template; (2) T7 RNA polymerase makes many copies of RNA: (3) a complementary DNA is made from the RNA by extension of a primer by reverse transcriptase; and (4) the RNA template is removed by ribonuclease H. This amplification method is useful for purposes such as genetic research and diagnostic assays.

REFERENCES:
patent: 4800159 (1992-06-01), Mullis et al.
patent: 5130238 (1992-07-01), Malek et al.
Kwoh et al., PNAS (U.S.A.) 86:1173-1177 (Feb., 1989).
Masukata et al., Cell 36: 513-522 (Feb., 1984).
Melton et al., Nuc. Acids Res. 12(18): 7035-7056 (1984).
Stoflet et al., Science 239: 491-494 (Jan. 29, 1988).
Krupp et al., Febs Letters 212(2): 271-275 (Feb. 1987).

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