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Publication Title | A PCR REACTOR WITH AN INTEGRATED ALUMINA ISOLATION MEMBRANE

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Supplementary Material (ESI) for Lab on a Chip This journal is (c) The Royal Society of Chemistry 2010

[May 04, 2010]

A PCR REACTOR WITH AN INTEGRATED ALUMINA ISOLATION MEMBRANE

Jitae Kima, Michael Mauka, Dafeng Chena, Xianbo Qiua, Jungkyu Kimb, Bruce Galeb, and Haim H. Baua,c

a. Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, Philadelphia, PA 19104-6315

b. Department of Mechanical Engineering, University of Utah, Salt Lake City, UT 84112

c. Corresponding author (bau@seas.upenn.edu)

ABSTRACT

Recently, there has been a growing interest in point of care devices capable of detecting

nucleic acids in clinical and environmental samples. Nucleic acid detection requires, however, various sample preparation steps that complicate device operation. An attractive remedy is to integrate many, if not all, sample preparation operations and nucleic acid amplification into a single reaction chamber. A microfluidic chip that integrates, in a single chamber, polymerase chain reaction (PCR) amplification with solid-phase extraction of nucleic acids using a nanoporous, aluminum oxide membrane (AOM) is described. Samples suspected of containing target bacteria and/or viruses are mixed with lysis agents and a chaotropic salt and loaded into a plastic chip housing a nanoporous, aluminum oxide membrane. The nucleic acids in the lysate bind to the membrane. The membrane is then washed, the chamber is filled with the PCR reaction reagents, and the chamber’s temperature is cycled to amplify the captured nucleic acids and produce detectable products. Both DNA and RNA (with reverse-transcription) isolation and amplification are demonstrated. Due to the dry membrane’s high resistance to liquid flow, a specialized flow control system was devised to facilitate sample introduction and membrane washing.

Keywords: Lab on a chip, Point of care, diagnostic device, PCR, RT, HIV, virus, bacteria

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