Novel DNA probes for sensitive DNA detection

Abstract

The ability to detect and interrogate DNA sequences allows further understanding and
diagnosis of genetic disease. The ability to perform such analysis of genetic material
requires highly selective and reliable technologies. Furthermore techniques which can use
simple and cheap equipment allow the use of such technologies for point of care analysis.

Described in this thesis are two novel DNA probe systems designed for mutation
discrimination and sequence recognition of PCR products. A homogenous PCR system
using HyBeacons® which utilise FRET to produce a three probe multiplex system and
surface enhanced Raman detection method. Both of these systems allow multiplex
detection of PCR products and mutation discrimination by melting temperature analysis.
The research reported includes investigations into the effects of different modifications to
improve the performance of HyBeacon® probes as well as the effect of different dyes in a
FRET system, including unique changes in the optical properties of such dyes. Also a
novel method of performing melting temperature analysis using an electrochemical
potential is reported.

In addition to the detection methods described this thesis includes initial work into the
stabilisation of quantum dot nanoparticles for their use in aqueous systems as a potential
alternative to fluorescent organic molecules.

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