Fluorerscence polarization detection of nucleic acid amplication

Details Fluorerscence polarization detection of nucleic acid amplication Fluorerscence polarization detection of nucleic acid amplication

1994-09-23

1997-01-14

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 935 78, C12P 1934, C12Q 168

Patent

active

055938671

ABSTRACT:
Fluorescence polarization methods for detection of nucleic acid amplification. A fluorescently labelled oligodeoxynucleotide probe is converted from single- to double-stranded form in a target dependent manner during amplification of the target sequence. This conformational change is accompanied by an increase in fluorescence polarization values. The increase in fluorescence polarization can be measured on a transient-state fluorometer in real-time during the amplification reaction without any physical manipulation of the sample. The inventive methods therefore provide a closed, homogeneous system for both amplification of target sequences and detection of amplification. Alternatively, amplification may be detected in the fluorometer after the amplification reaction is completed.

REFERENCES:
patent: 5270184 (1993-12-01), Walker et al.
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R. Devlin, et al. "Homogeneous detection of nucleic acids by transient-state polarized fluorescence" Clin. Chem. 39, 1939-1943 (1993).
G. T. Walker, et al. "Multiplex strand displacement amplification (SDA) and detection of DNA sequences from Mycobacterium tuberculosis and other mycobacteria" Nucl. Acids Res. 22, 2670-2677 (1994).
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J. C. Guatelli, et al. "Isothermal, in vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication" Proc. Natl. Acad. Sci. USA 87, 1874-1878 (1990).

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