Novel DNA probes for sensitive DNA detection

The ability to detect and interrogate DNA sequences allows further understanding and<br/>diagnosis of genetic disease. The ability to perform such analysis of genetic material<br/>requires highly selective and reliable technologies. Furthermore techniques which can use<br/>simple and cheap equipment allow the use of such technologies for point of care analysis.<br/><br/>Described in this thesis are two novel DNA probe systems designed for mutation<br/>discrimination and sequence recognition of PCR products. A homogenous PCR system<br/>using HyBeacons® which utilise FRET to produce a three probe multiplex system and<br/>surface enhanced Raman detection method. Both of these systems allow multiplex<br/>detection of PCR products and mutation discrimination by melting temperature analysis.<br/>The research reported includes investigations into the effects of different modifications to<br/>improve the performance of HyBeacon® probes as well as the effect of different dyes in a<br/>FRET system, including unique changes in the optical properties of such dyes. Also a<br/>novel method of performing melting temperature analysis using an electrochemical<br/>potential is reported.<br/><br/>In addition to the detection methods described this thesis includes initial work into the<br/>stabilisation of quantum dot nanoparticles for their use in aqueous systems as a potential<br/>alternative to fluorescent organic molecules.

Name dc:type.qualificationname

Ph.D.

Level dc:type.qualificationlevel

doctoral

Grantor dc:publisher.institution

University of Southampton

Year dc:date.issued

2010

Author dc:creator

• Richardson, James Alistair

Advisor dc:contributor.advisor

• Brown, Tom