Abstract
Simple, low-cost mutation detection assays that are suitable for low-throughput analysis are essential for diagnostic applications where the causative mutation may be different in every family. The mismatch oxidation assay is a simple optical absorbance assay to detect nucleotide substitutions, insertions, and deletions in heteroduplex DNA. The method relies on detecting the oxidative modification products of mismatched thymine and cytosine bases by potassium permanganate as it is reduced to manganese dioxide. This approach, unlike other methods commonly used to detect sequence variants, does not require costly labeled probes or primers, toxic chemicals, or a time-consuming electrophoretic separation step. The oxidation rate, and hence the presence of a sequence variant, is detected by measuring the formation of the potassium permanganate reduction product (hypomanganate diester), which absorbs at the 420-nm visible wavelength, using a standard UV/vis microplate reader.
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Acknowledgements
The authors wish to a thank Vincent Sesto for technical assistance with preparation of this manuscript and Sean Stockwell for critically reviewing the manuscript. This work was supported by an NH&MRC grant (to R.G.H.C.).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC 2003
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Tabone, T., Sallmann, G., Cotton, R.G.H. (2009). Mismatch Oxidation Assay: Detection of DNA Mutations Using a Standard UV/Vis Microplate Reader. In: Komar, A. (eds) Single Nucleotide Polymorphisms. Methods in Molecular Biology™, vol 578. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60327-411-1_15
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DOI: https://doi.org/10.1007/978-1-60327-411-1_15
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