Integrated polymerase chain reaction/electrophoresis instrument

Chemistry: electrical and wave energy – Apparatus – Electrophoretic or electro-osmotic apparatus

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Details

204450, 204451, 204452, 204600, 204603, 4352872, 4352885, G01N 2726, G01N 27447

Patent

active

061268046

ABSTRACT:
A new approach and instrument for field identification of micro-organisms and DNA fragments using a small and disposable device containing integrated polymerase chain reaction (PCR) enzymatic reaction wells, attached capillary electrophoresis (CE) channels, detectors, and read-out all on/in a small hand-held package. The analysis instrument may be made inexpensively, for example, of plastic, and thus is disposable, which minimizes cross contamination and the potential for false positive identification between samples. In addition, it is designed for multiple users with individual applications. The integrated PCR/CE is manufactured by the PCR well and CE channels are "stamped" into plastic depressions where conductive coatings are made in the wells and ends of the CE microchannels to carry voltage and current to heat the PCR reaction mixtures and simultaneously draw DNA bands up the CE channels. Light is transmitted through the instrument at appropriate points and detects PCR bands and identifies DNA fragments by size (retention time) and quantifies each by the amount of light generated as each phototransistor positioned below each CE channel detects a passing band. The instrument is so compact that at least 100 PCR/CE reactions/analyses can be performed easily on one detection device.

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A.T. Woolley et al "Functional Integration of PCR Amplification and Capillary Electrophoresis in a Microfabricated DNA Analysis Device" Analytical Chemistry, vol. 68, No. 23 (Dec. 1, 1996) 4081-4086.
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M.A. Burns "Microfabricated structures for integrated DNA analysis" Proceedings of the National Academy of Science, USA vol. 93 (May 1996) 5556-596.

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