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Title: DNA Hybridisation Affinity Chromatography: A Novel Method for Genetic Diagnostics
Author: Morris, John
ISNI:       0000 0001 2411 6591
Awarding Body: Newcastle University
Current Institution: University of Newcastle upon Tyne
Date of Award: 2008
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A novel method for separating DNA strands by sequence recognition is described. The procedure has potential benefits over other current methodologies in simplicity, economy and specificity in separating DNA single strands ofsimilar sequence. The method allows DNA strands within a sample the opportunity to hybridise multiple times with complementary probe DNA. The cumulative time spent in association in a stringent environment is a precise measure of the degree of complementarity between the probe and the DNA strands in the sample. This cumulative measurement is facilitated by moving the sample in a flow past probes that are tethered in a channel analogous with a chromatographic column. Under the right conditions of temperature and stringency sample DNA strands will associate transiently with stationary probes and be retained momentarily. The cumulative delay after many such encounters is a function of the affinity of the two matched, or closely matched DNA sequences. Distinguishing specific sequences of DNA, particularly in the presence of similar, but non-identical sequences, has broad application in clinical diagnostics and beyond. A high density of probes needs to be immobilised within the channel. Immobilisation of DNA probes to a gold-coated surface using gold-thiol covalent bonding was investigated but dropped in favour of a porous silica microparticle substrate with a larger surface area and less interference. The aim of demonstrating the process in its simplest, most economical form was met with disappointment; the relatively slight signal of increased retention time due to multiple, transient sequence-specific interactions was not consistently resolved through the broad imprecision of the chosen apparatus.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available