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Detection of a few DNA copies by real-time electrochemical polymerase chain reaction

Abstract

In the present work, accurate quantification over 10 to 108 DNA copies has been for the first time successfully achieved by real-time electrochemical PCR. This has been made possible thanks to the combine use of a fully automated house-built electrochemical qPCR device, optimized for parallel heating and electrochemical monitoring of up to 48 PCR solutions, and the appropriate selection of a DNA intercalating redox probe retaining a strong affinity binding to ds-DNA at the PCR measurement temperature of 72°C (corresponding to the PCR elongation step). This has also been achieved through the identification of the key parameters governing the onset electrochemical signal decrease and amplitude signal decrease as a function of the PCR cycle for a given DNA intercalating redox probe, allowing thus to predict the electrochemical PCR kinetic plots from the values of DNA affinity binding constant determined as a function of temperature. To the best of our knowledge, the analytical performances of the present electrochemical qPCR outperform all of that previously published, being even in terms of detection limit, dynamic range, reproducibility and melting curve analysis, competitive to that achieved on commercialized bench-top fluorescence-based qPCR instrument.

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Supplementary files

Publication details

The article was received on 12 Jun 2017, accepted on 28 Jul 2017 and first published on 28 Jul 2017


Article type: Paper
DOI: 10.1039/C7AN00978J
Citation: Analyst, 2017, Accepted Manuscript
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    Detection of a few DNA copies by real-time electrochemical polymerase chain reaction

    M. Moreau, S. Delile, A. Sharma, C. Fave, A. Perrier, B. Limoges and D. Marchal, Analyst, 2017, Accepted Manuscript , DOI: 10.1039/C7AN00978J

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