Detection of nucleic acids by fluorescence quenching

Details Detection of nucleic acids by fluorescence quenching Detection of nucleic acids by fluorescence quenching

1999-01-22

2000-10-10

Horlick, Kenneth R.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 911, 435 912, 536 221, 536 243, 536 2433, C12Q 168, C12P 1934, C07H 2104

Patent

active

061300470

ABSTRACT:
Detector nucleic acids are employed for detection of nucleic acid target sequences by fluorescence quenching mechanisms. The detector nucleic acid comprises at least two oligonucleotides and is partially single-stranded and partially double-stranded. One of the two dyes of a donor/acceptor dye pair is linked to the first oligonucleotide and the other is linked to a second oligonucleotide such that they are in close spatial proximity when the first and second oligonucleotides are base-paired and donor fluorescence is quenched. A single second oligonucleotide may be hybridized to the first oligonucleotide or multiple second oligonucleotides may be hybridized to the first oligonucleotide and to each other, forming a junction structure comprising multiple donor/acceptor dye pairs. The detector oligonucleotide retains its partially single-stranded and partially double-stranded conformation in the absence of target. In the presence of target, however, the second oligonucleotide(s) of the detector nucleic acid is/are completely or partially displaced from the first, increasing the distance between the donor and acceptor dyes and causing a change in fluorescence which may be detected as an indication of the presence of the target sequence.

REFERENCES:
patent: 5270184 (1993-12-01), Walker et al.
patent: 5348853 (1994-09-01), Wang et al.
patent: 5451503 (1995-09-01), Hogan et al.
patent: 5455166 (1995-10-01), Walker
patent: 5514546 (1996-05-01), Kool
patent: 5547861 (1996-08-01), Nadeau et al.
patent: 5607834 (1997-03-01), Bagwell
patent: 5627032 (1997-05-01), Ulanovsky
patent: 5691145 (1997-11-01), Pitner et al.
patent: 5712386 (1998-01-01), Wang et al.
M. S. Ellwood, et al. "Strand Displacement Applied to Assays with Nucleic Acid Probes" Clin. Chem. 32:1631-1636 (1986).
S. P. Lee et al. "A Fluorometric Assay for DNA Cleavage Reactions Characterized with BamHI Restriction Endonuclease" Anal. Biochem. 220:377-383 (1994).
L. G. Lee, et al. "Allelic discrimination by nick-translation PCR with fluorogenic probes" Acids Res. 21:3761-3766 (1993).
S. S. Ghosh, et al. "Real time kinetics of restriction endonuclease cleavage monitored by flourescence resonance energy transfer" Nucl. Acids Res. 22:3155-3159 (1994).
L. E. Morrison "Detection of Energy Transfer and Fluroescence Quenching" Nonisotopic DNA Prove Techniques, pp. 311-352 (1992).
P. S. Eis and D. P. Millar "Conformational Distributions of a Four-Way DNA Junction Revealed by Time-Resolved Fluorescence Resonance Energy Transfer" Biochem. 32:13852-13860 (1993).
M. Yang and D. P. Millar "Conformational Flexibility of Three-Way DNA Junctions Containing Unpaired Nucleotides" Biochem. 35:7959-7967 (1996).
R. M. Clegg, et al. "The Solution Structure of the Four-Way DNA Junction at Low-Salt Conditions: A Fluorescence Resonance Energy Transfer Analysis" Biophys. J. 66:99-109 (1994).
R. M. Clegg, et al. "The four-way DNA junction: A fluorescence resonance energy transfer study" Braz. J. Med. Biol. Res. 26:405-416 (1993).
R. M. Clegg, et al. "Fluorescence Resonance Energy Transfer Analysis of the Structure of the Four-Way DNA Junction" Biochem. 31:4846-4856 (1992).
D. M. J. Lilley and R. M. Clegg "The Structure of the Four-Way Junction in DNA" Annu. Rev. Biophys. Biomol. Sruct. 22:299-328 (1993).
J. E. Ladbury, et al. "The Thermodynamics of Formation of a Three-Strand, DNA Three-Way Junction Complex" Biochem. 33:6828-6833 (1994).
Y. Wang, et al. "Assembly and Characterization of Five-Arm and Six-Arm DNA Branched Junctions" Biochem. 30:5667-5674 (1991).
J. L. Kadrmas, et al. "Relative stabulities of DNA three-way, four-way and five-way junctions (multi-helix junction loops): unpaired nucleotides can be stabilizing or destabilizing" Nucl. Acids Res. 23:2212-2222 (1995).

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