Fluorescence polarization detection of nucleic acids

Details Fluorescence polarization detection of nucleic acids Fluorescence polarization detection of nucleic acids

1995-11-15

1997-06-24

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 435 9153, 536 243, 536 2433, 536 2532, 935 77, 935 78, C12P 1934, C12Q 168

Patent

active

056416334

ABSTRACT:
The present invention provides methods for detecting amplified or unamplified nucleic acid target sequences at increased temperatures by changes in fluorescence polarization. The decrease in fluorescence polarization associated with hybridization of oligonucleotides at higher, more stringent, temperatures is overcome by including a double-stranded DNA binding protein in the assay. At elevated temperatures, the double-stranded DNA binding protein restores, and often enhances, the magnitude of the change in fluorescence polarization associated with single- to double-stranded conversion of an oligonucleotide probe or primer.

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Cook et al., Analytical Biochemistry 190:331-330 1990.
Walker et al., Nuc. Acids Res. 24(2):348-353 Jan. 15, 1996.
G. T. Walker, et al. "Strand Displacement Amplification --an isothermal, in vitro DNA amplification technique" Nucl. Acids Res. 20:1691-1696 (1992).
G. T. Walker, et al. "Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system" Proc. Natl. Acad. Sci. USA 89:392-296 (1992).
A. Murakami, et al. "Fluorescent-labeled oligonucleotide probes: detection of hybrid formation in solution by fluorescence polarization spectroscopy" Nucl. Acids Res. 19:4097-4102 (1991).

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