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A label-free, surface-enhanced Raman spectroscopy-based assay for detecting DNA hybridization at an electrode surface and for distinguishing between mutations in DNA is demonstrated. Surface-immobilized DNA is exposed to a binding agent that is selective for dsDNA and acts as a reporter molecule. Upon application of a negative potential, the dsDNA denatures into its constituent strands, and the changes in the spectra of the reporter molecule are monitored. This method has been used to distinguish between a wild-type, 1653C/T single-point mutation and ΔF508 triplet deletion in the CFTR gene. The use of dsDNA-selective binding agents as reporter molecules in a discrimination assay removes the burden of synthetically modifying the target to be detected, while retaining flexibility in the choice of the reporter molecule.

Original publication

DOI

10.1021/ja304663t

Type

Journal article

Journal

J Am Chem Soc

Publication Date

29/08/2012

Volume

134

Pages

14099 - 14107

Keywords

Base Sequence, Biosensing Techniques, Coloring Agents, Cystic Fibrosis Transmembrane Conductance Regulator, DNA, DNA Mutational Analysis, Immobilized Nucleic Acids, Intercalating Agents, Nucleic Acid Hybridization, Point Mutation, Spectrum Analysis, Raman