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Towards multi-colour strategies for the detection of nucleic acid hybridization using quantum dots as energy donors in fluorescence resonance energy transfer (FRET)

Posted on:2007-08-20Degree:M.ScType:Thesis
University:University of Toronto (Canada)Candidate:Algar, Walter RussellFull Text:PDF
GTID:2441390005974740Subject:Chemistry
Abstract/Summary:
The unique optical properties of quantum dots (QDs) are of interest in the development of nucleic acid diagnostics. The potential for a simultaneous two-colour diagnostic scheme for nucleic acids operating on the basis of fluorescence resonance energy transfer (FRET) has been demonstrated. Upon ultraviolet excitation, two-colours of CdSe/ZnS quantum dots with conjugated oligonucleotide probes acted as energy donors yielding FRET-sensitized acceptor emission upon hybridization with fluorophore labeled target oligonucleotides. The use of an intercalating dye to improve signal-to-noise was also demonstrated. The major limitation of the system was the non-specific adsorption of oligonucleotides, which was characterized extensively. Adsorptive interactions were found to affect the conformation of oligonucleotides conjugated to QDs, the kinetics of hybridization with QD-DNA conjugates, and the thermal stability of those hybrids. In addition, it was found that thiol-alkyl-acid capped QDs exhibited pKa correlated ligand-chromism and radiative decay rate-driven changes in quantum yield.
Keywords/Search Tags:Quantum, Nucleic, Energy, Qds, Hybridization
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