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Electronic Detection of Single-Base Mismatches in DNA with Ferrocene-Modified Probes

Yu, C. J. and Wan, Yanjian and Yowanto, Handy and Li, Jie and Tao, Chunlin and James, Matthew D. and Tan, Christine L. and Blackburn, Gary F. and Meade, Thomas J. (2001) Electronic Detection of Single-Base Mismatches in DNA with Ferrocene-Modified Probes. Journal of the American Chemical Society, 123 (45). pp. 11155-11161. ISSN 0002-7863. https://resolver.caltech.edu/CaltechAUTHORS:20170405-114813362

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Abstract

Genotyping and gene-expression monitoring is critical to the study of the association between genetics and drug response (pharmacogenomics) and the association of sequence variation with heritable phenotypes. Recently, we developed an entirely electronic method for the detection of DNA hybridization events by the site-specific incorporation of ferrocenyl derivatives into DNA oligonucleotides. To perform rapid and accurate point mutation detection employing this methodology, two types of metal-containing signaling probes with varying redox potentials are required. In this report we describe a new ferrocene-containing phosphoramidite 9 that provides a range of detectable redox potentials. Using automated DNA/RNA synthesis techniques the two ferrocenyl complexes were inserted at various positions along oligonucleotide probes. Thermal stability analysis of these metal-containing DNA oligonucleotides indicates that incorporation of 9 resulted in no destabilization of the duplex. A mixture of oligonucleotides containing compounds 9 and I was analyzed by alternating current voltammetry (ACV) monitored at the 1st harmonic. The data demonstrate that the two ferrocenyl oligonucleotide derivatives can be distinguished electrochemically. A CMS-DNA array was prepared on an array of gold electrodes on a printed circuit board substrate with a self-assembled mixed monolayer, coupled to an electronic detection system. Experiments for the detection of a single-base match utilizing two signaling probes were carried out. The results demonstrate that rapid and accurate detection of a single-base mismatch can be achieved by using these dual-signaling probes on CMS-DNA chips.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja010045fDOIArticle
http://pubs.acs.org/doi/full/10.1021/ja010045fPublisherArticle
Additional Information:© 2001 American Chemical Society. Received 4 January 2001. Published online 23 October 2001. Published in print 1 November 2001. We wish to thank Prof. Yitzhak Tor for helpful discussions during the preparation of this manuscript and Dr. Jost Viemetter for acquiring T_m data. We are indebted to the DNA-Chip production team at Motorola Clinical MicroSensors for preparing the chips used in this study.
Issue or Number:45
Record Number:CaltechAUTHORS:20170405-114813362
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170405-114813362
Official Citation:Electronic Detection of Single-Base Mismatches in DNA with Ferrocene-Modified Probes C. J. Yu, Yanjian Wan, Handy Yowanto, Jie Li, Chunlin Tao, Matthew D. James, Christine L. Tan, Gary F. Blackburn, and Thomas J. Meade Journal of the American Chemical Society 2001 123 (45), 11155-11161 DOI: 10.1021/ja010045f
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:75737
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:05 Apr 2017 18:58
Last Modified:03 Oct 2019 16:53

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