Please use this identifier to cite or link to this item: http://hdl.handle.net/10397/27028
Title: Electrochemistry-based real-time PCR on a microchip
Authors: Yeung, SSW
Lee, TMH 
Hsing, IM
Issue Date: 2008
Publisher: American Chemical Society
Source: Analytical chemistry, 2008, v. 80, no. 2, p. 363-368 How to cite?
Journal: Analytical chemistry 
Abstract: The development of handheld instruments for point-of-care DNA analysis can potentially contribute to the medical diagnostics and environmental monitoring for decentralized applications. In this work, we demonstrate the implementation of a recently developed electrochemical real-time polymerase chain reaction (ERT-PCR) technique on a silicon-glass microchip for simultaneous DNA amplification and detection. This on-chip ERT-PCR process requires the extension of an oligonucleotide in both solution and at solid phases and intermittent electrochemical signal measurement in the presence of all the PCR reagents. Several important parameters, related to the surface passivation and electrochemical scanning of working electrodes, were investigated. It was found that the ERT-PCR's onset thermal cycle (∼3-5), where the analytical signal begins to be distinguishable from the background, is much lower than that of the fluorescence-based counterparts for high template DNA situations (3 × 106 copies/μL). By carefully controlling the concentrations of the immobilized probe and the enzyme polymerase, improvements have been made in obtaining a meaningful electrochemical signal using a lower initial template concentration. This ERT-PCR technique on a microchip platform holds significant promise for rapid DNA detection for point-of-care testing applications.
URI: http://hdl.handle.net/10397/27028
ISSN: 0003-2700
EISSN: 1520-6882
DOI: 10.1021/ac071198+
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