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Issue 2, 2003
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Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

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Abstract

Microfabricated silicon/glass-based devices with functionalities of simultaneous polymerase chain reaction (PCR) target amplification and sequence-specific electrochemical (EC) detection have been successfully developed. The microchip-based device has a reaction chamber (volume of 8 µl) formed in a silicon substrate sealed by bonding to a glass substrate. Electrode materials such as gold and indium tin oxide (ITO) were patterned on the glass substrate and served as EC detection platforms where DNA probes were immobilized. Platinum temperature sensors and heaters were patterned on top of the silicon substrate for real-time, precise and rapid thermal cycling of the reaction chamber as well as for efficient target amplification by PCR. DNA analyses in the integrated PCR-EC microchip start with the asymmetric PCR amplification to produce single-stranded target amplicons, followed by immediate sequence-specific recognition of the PCR product as they hybridize to the probe-modified electrode. Two electrochemistry-based detection techniques including metal complex intercalators and nanogold particles are employed in the microdevice to achieve a sensitive detection of target DNA analytes. With the integrated PCR-EC microdevice, the detection of trace amounts of target DNA (as few as several hundred copies) is demonstrated. The ability to perform DNA amplification and EC sequence-specific product detection simultaneously in a single reaction chamber is a great leap towards the realization of a truly portable and integrated DNA analysis system.

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Publication details

The article was received on 20 Jan 2003, accepted on 25 Mar 2003 and first published on 17 Apr 2003


Article type: Paper
DOI: 10.1039/B300799E
Citation: Lab Chip, 2003,3, 100-105
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    Microfabricated PCR-electrochemical device for simultaneous DNA amplification and detection

    T. M. Lee, M. C. Carles and I. Hsing, Lab Chip, 2003, 3, 100
    DOI: 10.1039/B300799E

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