An optimal design method for preventing air bubbles in high-temperature microfluidic devices |
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Authors: | Tsuyoshi Nakayama Ha Minh Hiep Satoshi Furui Yuji Yonezawa Masato Saito Yuzuru Takamura Eiichi Tamiya |
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Institution: | (1) School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1292, Japan;(2) Department of Applied Physics, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan;(3) National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan;(4) Industrial Research Institute of Ishikawa, 2-1 Kuratsuki, Kanazawa, Ishikawa 920-8203, Japan; |
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Abstract: | DNA analysis with the polymerase chain reaction (PCR) has become a routine part of medical diagnostics, environmental inspections,
food evaluations, and biological studies. Furthermore, the development of a microscale PCR chip is an essential component
of studies aimed at integrating PCR into a micro total analysis system (μ-TAS). However, the occurrence of air bubbles in
microchannels complicates this process. In this study, we investigated a new technique based on the fluid dynamics of laminar
flow that utilizes a small amount of mineral oil at the beginning of sample injection to prevent air bubbles from occurring
in microchannels. We also further optimized the pressure, the length of the pressurizing channel and the volume of oil, thus
making our microfluidic device more useful for high-temperature PCR. Additionally, quantitative continuous-flow PCR was performed
using the optimized PCR chip in order to detect genetically modified (GM) maize. DNA was extracted from GM maize, MON 810,
and non-GM maize at several concentrations from 0% (w/v) to 100% (w/v). The DNA amplification signals were then analyzed on
the PCR chip using a laser-based system. The signal from our microfluidic PCR chip was found to increase in direct proportion
to the initial GM maize concentration. |
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