Detection of single nucleotide polymorphisms using a DNA Holliday junction nanoswitch?a high-throughput fluorescence lifetime assay

McGuinness, Colin D., Nishimura, Mira K. Y., Keszenman-Pereyra, David, Dickinson, Paul, Campbell, Colin J., Bachmann, Till T., Ghazal, Peter ORCID: https://orcid.org/0000-0003-0035-2228 and Crain, Jason 2009. Detection of single nucleotide polymorphisms using a DNA Holliday junction nanoswitch?a high-throughput fluorescence lifetime assay. Molecular BioSystems 6 (2) , pp. 386-390. 10.1039/B913455G

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Abstract

We report a simple DNA sensor device, using a combination of binding and conformational switching, capable of rapid detection of specific single nucleotide polymorphisms in an unlabelled nucleic acid target sequence. The detection is demonstrated using fluorescence lifetime measurements in a high-throughput micro plate reader instrument based on the time-correlated single-photon counting technique. The sensor design and instrumental architecture are capable of detecting perturbations in the molecular structure of the probe–target complex (which is similar to that of a Holliday junction), due to a single base pair mismatch in a synthetic target. Structural information, including fluorophore separations, is obtained using time-resolved Förster resonance energy transfer between two fluorophores covalently bound to the probe molecule. The two probes required are designed to detect a single nucleotide polymorphism from a sequence present on each of the two copies of human chromosome 11

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Publisher:	Royal Society of Chemistry
ISSN:	1742-206X
Date of Acceptance:	21 September 2009
Last Modified:	23 Oct 2022 14:16
URI:	https://orca.cardiff.ac.uk/id/eprint/113222

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