Abstract
We report a straightforward protocol for the detection of mutated DNA extracted from cancer cells. The assay combines a step-wise solid-phase hybridization and a readout by fluorescence emission. We detect a single-nucleotide polymorphism in two human oncogenes, BRAF and EGFR, and reach a limit of the detection of 300 pM by conventional fluorometry. The protocol described herein may be used as a foundation for development of automatic optimized assays capable for detection of mutant DNA and RNA in vitro and in cells.
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Acknowledgments
The work is supported by Villum Foundation Young Investigator Programme, award number 13152.
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Taskova, M., Astakhova, K. (2020). Solid-Phase Hybridization Assay for Detection of Mutated Cancer DNA by Fluorescence. In: Astakhova, K., Bukhari, S. (eds) Nucleic Acid Detection and Structural Investigations. Methods in Molecular Biology, vol 2063. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0138-9_4
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DOI: https://doi.org/10.1007/978-1-0716-0138-9_4
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0138-9
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