| Summary: | 碩士 === 國立成功大學 === 工程科學系碩博士班 === 96 === A new micromachined circulating polymerase chain reaction (PCR) chip is reported in this study. A new suction-type liquid transportation mechanism and three microvalves were used to comprise a new microfluidic control module to rapidly transport the DNA samples and PCR reagents around three bio-reactors operating at three different temperatures. When operating at a frequency of 14.29 Hz and a pressure of 3 psi, the sample flow rate of the microfluidic control module can be as high as 18 μL/sec. In addition, an array-type microheaters and open-type reaction chambers were adopted to improve the thermal uniformity in the reaction chambers and to facilitate the operation for temperature calibration. Experimental data from infrared images showed that the percentage of the uniformity area inside the reaction chamber with a thermal variation less than 1℃�awere over 90% for a denaturing temperature of 94℃. Three array-type heaters and temperature sensors were integrated into this new circulating PCR chip to modulate three specific operating temperatures for denaturation, annealing, and extension of nucleic acids. With this approach, the cycle numbers and reaction time of three reaction steps for a PCR procedure can be individually adjusted by the microfluidic control module. To verify the performance of this circulating PCR chip, a PCR process to amplify a detection gene (150 base pairs) associated with hepatitis C virus was performed. Experimental results showed that biosamples with concentrations ranging from 108 to 5×10^3 copies/mL can be successfully amplified by using this PCR chip. Therefore, the new circulating PCR chip can provide a useful platform for genetic identification and diagnosis.
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