| Summary: | 碩士 === 國立臺灣大學 === 應用力學研究所 === 93 === This thesis presents a new device to perform polymerase chain reaction (PCR) using polydimethylsiloxane (PDMS) polymer as disposable PCR channel. Particular feature in the device include gravity-fed, circularly reciprocating action to ensure the PCR reagent is in contact with a specific temperature region during the PCR cycle of three temperatures – 94oC, 55oC, 72oC. The PCR channel (3.5 mm width by 1 mm height) is fabricated by molding PDMS against a laser scriber engraved PMMA master, with a 4 µm thick parylene film coated on the PCR channel inner and outer surfaces to alleviate bubble formation at high temperature. Simulation, using ANSYS, of heat transfer of the device is also undertaken to aid the design of the device.
Results from temperature measurement, using resistive temperature detectors (RTD) sensors, show that the reagent can be cooled down to 55oC (from 94oC ) in 30 sec, or heated up to 72oC and 94oC in 15 sec, in agreement with ANSYS predictions. PCR is demonstrated by a 773 bps DNA template, which is incubated at 94oC for 30 sec, 55oC for 1 min., and 72oC for 1 min. to perform DNA denaturation, primer annealing, and extension of DNA template, respectively. Using the present first-generation device, DNA amplification is clearly proven, however, the efficiency is not as good as commercial PCR machine. Evaporation in the reagent is believed to be the key problem that needs to be improved.
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