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Amplified Analysis of DNA by the Autonomous Assembly of Polymers Consisting of DNAzyme Wires

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Institute of Chemistry, The Hebrew University of Jerusalem, Jerusalem 91904, Israel
Cite this: J. Am. Chem. Soc. 2011, 133, 43, 17149–17151
Publication Date (Web):September 28, 2011
https://doi.org/10.1021/ja2076789
Copyright © 2011 American Chemical Society

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    Abstract

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    A systematic study of the amplified optical detection of DNA by Mg2+-dependent DNAzyme subunits is described. The use of two DNAzyme subunits and the respective fluorophore/quencher-modified substrate allows the detection of the target DNA with a sensitivity corresponding to 1 × 10–9 M. The use of two functional hairpin structures that include the DNAzyme subunits in a caged, inactive configuration leads, in the presence of the target DNA, to the opening of one of the hairpins and to the activation of an autonomous cross-opening process of the two hairpins, which affords polymer DNA wires consisting of the Mg2+-dependent DNAzyme subunits. This amplification paradigm leads to the analysis of the target DNA with a sensitivity corresponding to 1 × 10–14 M. The amplification mixture composed of the two hairpins can be implemented as a versatile sensing platform for analyzing any gene in the presence of the appropriate hairpin probe. This is exemplified with the detection of the BRCA1 oncogene.

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    DNA sequences, experimental procedures and methods, results for analysis of target (4) by the Mg2+-dependent DNAzyme subunits (1) and (2), and discussion of the amplified analysis of target (4) using functional hairpins (5) and (9) via “one-sided” DNAzyme nanowires. This material is available free of charge via the Internet at http://pubs.acs.org.

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