Use this URL to cite or link to this record in EThOS: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.551559
Title: Systematic quantitative analysis of the comet assay
Author: McArt, Darragh Gerrard
Awarding Body: University of Ulster
Current Institution: Ulster University
Date of Award: 2010
Availability of Full Text:
Access from EThOS:
Abstract:
The single cell gel electrophoresis assay (comet assay) has been widely used over the last 30 years. It is a robust method of estimating DNA movement under perturbation. In the assay cells are embedded in agarose on a frosted slide. The amount of broken or fragmented DNA that is removed from the central body, after the cell has been lysed and electrophoresed, is estimated as the total volume of DNA damage in the form of strand breaks caused by the insult. The damage is quantified by fluorescence microscopy with analysis software. It is a simple technique that is efficient and relatively quick. With the comet methodology there is much debate on the underlying mechanisms of cellular damage and repair and over how strongly the comet shape reflects the DNA damage that has occurred. It is often used as a quick-fire detection method to back up other analysis methods with very little thought given to the information it provides. The technique itself is open to experimenter bias at the microscopy level, where researchers have the freedom to select which comets to analyse. Addressing these issues along with those of standardising the technique and developing novel methods of comet analysis may enhance the information it provides. Using statistical and computational methods, it is proposed that a simulation of the comet would provide important information about DNA damage at a cellular level while offering impartial judgement and reproducibility. Information is available on some aspects of comet structure in published literature and with new techniques both computationally and experimentally it has become possible to examine intrinsic information on comet structure. Information was also established via novel and existing laboratory techniques and also through microscopy techniques to examine architecture. A computational modelling and simulation approach can then be used to ask important biological questions on aspects of DNA damage and repair at a cellular level.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.551559  DOI: Not available
Share: