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Single-mismatch detection using gold-quenched fluorescent oligonucleotides

Nature Biotechnology volume 19, pages 365–370 (2001)Cite this article

An Erratum to this article was published on 01 July 2001

Abstract

Here we describe a hybrid material composed of a single-stranded DNA (ssDNA) molecule, a 1.4 nm diameter gold nanoparticle, and a fluorophore that is highly quenched by the nanoparticle through a distance-dependent process. The fluorescence of this hybrid molecule increases by a factor of as much as several thousand as it binds to a complementary ssDNA. We show that this composite molecule is a different type of molecular beacon with a sensitivity enhanced up to 100-fold. In competitive hybridization assays, the ability to detect single mismatch is eightfold greater with this probe than with other molecular beacons.

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Figure 1: Schematic drawings of the two conformations of the dye–oligonucleotide–gold conjugate and of the gold-quenched molecular beacon.
Figure 2: Gel electrophoresis of reaction products.
Figure 3: Efficiency of the quenching of gold nanoparticles.
Figure 4: Comparison of single-mismatch detection with gold-quenched beacons versus DABCYL-quenched beacons.

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Acknowledgements

We thank G. Bonnet for enlightening discussions and suggestions, and for his help with the fluorescence correlation spectroscopy measurements. We thank R. Bar-Ziv, G.V. Shivashankar, T. Tomoda, and A. Mehta for interesting discussions. The help of A. Roll-Mecak and D. Stettler was instrumental in the final writing of this manuscript. We also thank the reviewers for improvement to the manuscript. This research was supported by the Mathers Foundation.

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Author notes

  1. Michel Calame

    Present address: Institute of Physics, Klingelbergstrasse 82, CH-4056, Basel, Switzerland

Authors and Affiliations

  1. Center for Studies in Physics and Biology, The Rockefeller University, 1230 York Avenue, New York, 10021, NY

    Benoit Dubertret, Michel Calame & Albert J. Libchaber

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  1. Benoit Dubertret
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Correspondence to Benoit Dubertret.

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Dubertret, B., Calame, M. & Libchaber, A. Single-mismatch detection using gold-quenched fluorescent oligonucleotides. Nat Biotechnol 19, 365–370 (2001). https://doi.org/10.1038/86762

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