Document Type	Master's Dissertation
Author	Van Zuydam, Natalie Rachel
URN	etd-10222009-102934
Document Title	Identification of Leptographium species by oligonucleotide discrimination on a DNA microarray
Degree	MSc
Department	Genetics
Supervisor	Advisor Name Title Dr K Jacobs Co-Supervisor Prof M J Wingfield Co-Supervisor Prof B D Wingfield Supervisor
Keywords	Leptographium microarray species identification
Date	2009-09-02
Availability	unrestricted
Abstract Leptographium is an anamorph genus within the Ophiostomatoid group of fungi and represents a unique case for molecular applications. The genus has a near complete sequence data available for three genes across all known species. This characteristic makes it a perfect test group for investigating applications of new diagnostic techniques within ascomycetes. Probes and primers, for microarrays, are designed from phylogenetically useful gene regions and are fabricated onto a solid substrate using printing technology. The sample is prepared using PCR and is hybridised to the probes under stringent conditions. The resulting fluorescent pattern is rigorously analysed to distinguish species from each other. Diagnostic PCR uses primers that are designed in similar way to the way probes are designed for microarrays and indicate the presence of a species through positive amplification. This research methodology will be applied to Leptographium to evaluate the efficacy of microarray technology for discriminating species within that genus. The data gained from this research study will be used in applications for other genera using microarray technology. © 2009, University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. Please cite as follows: Van Zuydum, N 2009, Identification of Leptographium species by oligonucleotide discrimination on a DNA microarray, MSc dissertation, University of Pretoria, Pretoria, viewed yymmdd < http://upetd.up.ac.za/thesis/available/etd-10222009-102934 / > E1490/gm
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