Method for detection of nucleic acid targets by amplification an

Details Method for detection of nucleic acid targets by amplification an Method for detection of nucleic acid targets by amplification an

1995-11-15

1998-09-01

Arthur, Lisa B.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 9121, 435 912, C12Q 168, C12P 1934

Patent

active

058009892

ABSTRACT:
Fluorescence polarization methods for detection of nucleic acid amplification at thermophilic temperatures employ a fluorescently labeled oligonucleotide signal primer which is converted from single- to double-stranded form in a target amplification-dependent manner. This conformational change is accompanied by an increase in fluorescence polarization values. The decrease in FP typically observed for the duplex at elevated temperatures is overcome by double-stranded DNA binding proteins which are believed to stabilize the double-stranded structure by reducing the single-strandedness normally associated with higher temperatures. The inventive methods provide a closed, homogeneous system for amplification and detection of amplification in real-time or at an endpoint.

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Wright, D.J. et al. (May 1995) Abstracts 95th Gen. Mtg. Am. Soc. Microbiol. 95(0): 133.
G. T. Walker, et al. "Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system" Proc. Natl. Acad. Sci. USA 89, 392-396 (1992).
G. T. Walker, et al. "Strand displacement amplification--an isothermal, in vitro DNA amplification technique" Nucl. Acids Res. 20, 1691-1696 (1992).
A. Murakami, et al. "Fluorescent-labeled oligonucleotide probes: detection of ohybrid formation in solution by fluorescence polarization spectroscopy" Nucl. Acids Res. 19, 4097-4102 (1991).

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