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Continuous Flow Polymerase Chain Reaction in Low Temperature Co-Fired Ceramics
Keywords: PCR, LTCC, DNA
The increasing need for rapid detection of certain biological species, such as malaria or anthrax, requires the mobilization of the various processes involved. One particular process involved in the accurate/rapid detection of biological species is Polymerase Chain Reaction (PCR). PCR is a type of DNA amplification that involves a sample experiencing three precisely controlled temperature zones (<0.5°C variation) for a certain amount of time; the temperatures and dwell times of each zone varies with species. A Continuous Flow PCR device has been designed and fabricated using Low Temperature Co-Fired Ceramics (LTCC). LTCC is desirable due to its non-reactive properties, ability to create internal three dimensional structures, low cost, compatibility with embedded heaters, portability and reusability. This work will focus on a device that contains three temperature zones and a single continuous channel internally. To facilitate an ample result, the DNA must pass through these zones a total of 30-40 times resulting in over a billion copies of the original sample. The initial internal channel geometry and heater sizing were determined using an analytical and heat transfer model. The first zone was fabricated with the model results and tested with water to develop a control system, capable of precise thermal management. Results from the first zone were applied to the design of one complete cycle (3 zones) and then to an 8 cycle. The multiple cycle design utilizes a simple, radially inward, channel configuration, which allows for continuous flow of the sample. Embedded temperature sensors, whose impedance is a function of temperature, have been developed to monitor each zone. This data will be plotted and will show the control system's ability to maintain minimal temperature variation and the time to reach steady state. Additionally, thermal images of LTCC surface indicate the device's ability to thermally isolate each zone.
Andrew Vissotski, Undergraduate Research Assistant
Boise State University
Boise, ID

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