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PDFOpen Access (License Unspecified), Open Access
Thesis
Development and optimization of a quantitative DNA base excision assay
Master of Science (M.S.), Drexel University
Sep 2008
DOI:
https://doi.org/10.17918/etd-2914
Abstract
The base excision repair (BER) system routinely removes damaged bases from DNA to conserve the integrity of the genome. The aims of this study were to design and optimize an assay using fluorescent kinetic data from molecular beacon (MB) oligonucleotides to generate an enzymatic model of base excision. Damaged bases, including 8-oxo-7,8-dihydroguanine (8oxoG) and uracil, were incorporated within 5'-fluorescein, 3'-Dabcyl labeled stem-loop oligonucleotides for use in this repair assay. Fluorescence data was recorded to monitor the kinetics of the base excision. Parameters were estimated using nonlinear regression to characterize the efficiency of repair proteins in whole cell extracts. This assay provides a simple, reproducible, and quantitative method to determine BER efficiency as a means of characterizing DNA repair capacity in cells.
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Details
- Title
- Development and optimization of a quantitative DNA base excision assay
- Creators
- Christopher Arif Abdullah - DU
- Contributors
- Bahrad Ali Sokhansanj (Advisor) - Drexel University (1970-)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems; Drexel University
- Other Identifier
- 2914; 991014632049904721