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Mutation detection by electrocatalysis at DNA-modified electrodes

Boon, Elizabeth M. and Ceres, Donato M. and Drummond, Thomas G. and Hill, Michael G. and Barton, Jacqueline K. (2000) Mutation detection by electrocatalysis at DNA-modified electrodes. Nature Biotechnology, 18 (12). pp. 1096-1100. ISSN 1087-0156. doi:10.1038/80301.

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Detection of mutations and damaged DNA bases is important for the early diagnosis of genetic disease. Here we describe an electrocatalytic method for the detection of single-base mismatches as well as DNA base lesions in fully hybridized duplexes, based on charge transport through DNA films. Gold electrodes modified with preassembled DNA duplexes are used to monitor the electrocatalytic signal of methylene blue, a redox-active DNA intercalator, coupled to [Fe(CN)_6]^3-. The presence of mismatched or damaged DNA bases substantially diminishes the electrocatalytic signal. Because this assay is not a measure of differential hybridization, all single-base mismatches, including thermodynamically stable GT and GA mismatches, can be detected without stringent hybridization conditions. Furthermore, many common DNA lesions and "hot spot" mutations in the human p53 genome can be distinguished from perfect duplexes. Finally, we have demonstrated the application of this technology in a chip-based format. This system provides a sensitive method for probing the integrity of DNA sequences and a completely new approach to single-base mismatch detection.

Item Type:Article
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URLURL TypeDescription ReadCube access
Boon, Elizabeth M.0000-0003-1891-839X
Barton, Jacqueline K.0000-0001-9883-1600
Additional Information:© 2000 Nature Publishing Group. Received 14 June 2000; accepted 25 July 2000. We are grateful to the NIH for financial support (GM61077). We also thank M.J. Fitzsimmons and J.L. Lamb in Device Research at JPL for assistance in the fabrication of the DNA chips.
Funding AgencyGrant Number
Subject Keywords:DNA chip; DNA monolayer; SNPs; mutation detection; damage detection; electrochemistry; electrocatalysis; p53 gene; methylene blue; DNA-mediated electron transfer
Issue or Number:12
Record Number:CaltechAUTHORS:20150408-152315146
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Official Citation:Boon, E. M., Ceres, D. M., Drummond, T. G., Hill, M. G., & Barton, J. K. (2000). Mutation detection by electrocatalysis at DNA-modified electrodes. [10.1038/80301]. Nat Biotech, 18(10), 1096-1100.
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:56501
Deposited On:08 Apr 2015 22:34
Last Modified:10 Nov 2021 21:00

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