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Multicolor molecular beacons for allele discrimination
Nature Biotechnologyvolume 16, pages49–53 (1998)Cite this article
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Abstract
Molecular beacons are hairpin-shaped oligonucleotide probes that report the presence of specific nucleic acids in homogenous solutions. When they bind to their targets they undergo a conformationsl reorganization that restores the fluorescence of an internally quenched fluorophore. We found that their hairpin conformation enables the use of a wide variety of differently colored fluorophores. Using several molecular beacons, each designed to recognize a different target and each labeled with a different fluorophore, we demonstrate that multiple targets can be distinguished in the same solution, even if they differ from one another by as little as a single nucleotide. A comparison of “hairpin probes” with corresponding “linear probes” confirms that the presence of the hairpin stem in molecular beacons significantly enhances their specificity.
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References
Tyagi, S. and Kramer, F.R. 1996. Molecular beacons: probes that fluoresce upon hybridization.Bio/Technology14: 303–308.
Matsuo, T. 1998. In situ visualization of messenger RNA for basic fibroblast growth factor in living cells.Biochim. Biophys. Acts.1379. In press.
Matayoshi, E.D., Wang, G.T., Krafft, G.A., and Erickson, J. 1990. Novel fluoro-genic substrates for assaying retroviral proteases by resonance energy transfer.Science247: 954–958.
Stryer, L. 1978. Fluorescence energy transfer as a spectroscopic ruler.Ann. Rev. Biochem.47: 819–846.
Haugland, R.P., Yguerabide, J., and Stryer, L. 1969. Dependence of the kinetics of singlet-singlet energy transfer on spectral overlap.Proc. Natl. Acad. Sci. USA63: 23–30.
Dexter, D.L. 1953. A theory of sensitized luminescence in solids.J. Chem. Phys.21: 836–850.
Turro, N.J. 1991.Modern Molecular Photochemistry. University Science Books, Mill Valley, CA.
Ikuta, S., Takagi, K., Wallace, R.B., and Itakura, K. 1987. Dissociation kinetics of 19 base paired oligonucleotide-DNA duplexes containing different single mismatched base pairs.Nucl. Acids Res.15: 797–811.
Tibanyenda, N., De Bruin, S.H., Haasnoot, C.A.G., van der Marel, G.A. van Boom, J.H., and Hilbers, C.W. 1984. The effect of single base-pair mismatches on duplex stability of d(T-A-T-T-A-A-T-A-T-C-A-A-G-T-T-G) • (C-A-A-C-T-T-G-A-T-A-T-T-A-A-T-A).Eur. J. Biochem.139: 19–27.
Werntges, H., Steger, G., Riesner, D. and Fritz, H.J., 1986. Mismatches in DNA double strands: thermodynamic parameters and their correlation to repair efficiencies.Nucl, Acids Res.14: 3773–3790.
Kostrikis, L.G., Tyagi, S., Mhlanga, M.M., Ho, D.D., and Kramer, F.R. 1998. Spectral genotyping of human alleles.Science279. In press.
Giesendorf, B.A.J., Vet, J.A.M., Tyagi, S., Mensink, E.J.M.G., Trijbels, F.J.M., and Blom, H.J. 1998. Molecular beacons: a new approach for semi-automated mutation analysis.Clin. Chem.44. In press.
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Department of Molecular Genetics, Public Health Research Institute, 455 First Ave., New York, NY, 10016
Sanjay Tyagi, Diana P. Bratu & Fred Russell Kramer
- Sanjay Tyagi
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- Diana P. Bratu
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- Fred Russell Kramer
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Correspondence toSanjay Tyagi.
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Tyagi, S., Bratu, D. & Kramer, F. Multicolor molecular beacons for allele discrimination.Nat Biotechnol16, 49–53 (1998). https://doi.org/10.1038/nbt0198-49
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