Novel DNA probes for sensitive DNA detection

The ability to detect and interrogate DNA sequences allows further understanding anddiagnosis of genetic disease. The ability to perform such analysis of genetic materialrequires highly selective and reliable technologies. Furthermore techniques which can usesimple 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 mutationdiscrimination and sequence recognition of PCR products. A homogenous PCR systemusing HyBeacons® which utilise FRET to produce a three probe multiplex system andsurface enhanced Raman detection method. Both of these systems allow multiplexdetection of PCR products and mutation discrimination by melting temperature analysis.The research reported includes investigations into the effects of different modifications toimprove the performance of HyBeacon® probes as well as the effect of different dyes in aFRET system, including unique changes in the optical properties of such dyes. Also anovel method of performing melting temperature analysis using an electrochemicalpotential is reported.In addition to the detection methods described this thesis includes initial work into thestabilisation of quantum dot nanoparticles for their use in aqueous systems as a potentialalternative to fluorescent organic molecules