Development of a Novel Convective Polymerase Chain Reaction System with Single-Temperature Control
Date Issued
2007
Date
2007
Author(s)
Hsieh, Yi-Fan
DOI
zh-TW
Abstract
Development of a Novel Convective Polymerase Chain Reaction System with Single-Temperature Control
Hsieh, Yi-Fan
Department of Mechanical Engineering, National Taiwan University,
Taipei, Taiwan, R.O.C.
ABSTRACT
The concept of convective PCR (Convective Polymerase Chain Reaction) was revealed in 2002. The convection is generated by the temperature gradient controlled by a couple of heaters. This essay focuses on the improvement of the design of the heater – single heating source. We provided a glass capillary tube of 75μL used as the vessel, combined with constant temperature control in the oil-bath. The temperature of the bottom of the capillary tube is kept 94℃ for denaturing. The surface of the capillary tube is cooled by contacting the room temperature. Thus the convection observed is an effective method for amplification of the oligo-nucleic acid and the detection of the mutation and infected disease.
By using the particle image velocimetry, PIV method, to observe the convection in the tube, the flow and the velocity distribution are obtained. Meanwhile, the convection from the bottom to the top of the tube can be inspected to calculate the optimized parameters for PCR. Thermal couples and IR ( Infrared Spectroscopy ) are also employed for the inspection of the temperature distribution of the capillary tube. The simulated results calculated by using the commercial suite Fluent is also compared with the observed PIV results. Since the experimental results show that the 122-bp plasmid of hepatitis B virus of 30 copies/ tube can be specifically amplified successfully, we can conclude that the velocity of the fluid is strictly related to the probability of the successful PCR.
This research proves that the convection caused by a single heater can successfully execute PCR. Due to the conclusions of this essay, we believe that it is helpful to establish the real-time convective PCR.
Subjects
熱對流
聚合酶
連鎖反應
紅外線測溫儀
粒子影像測速儀
Convection
Polymerase Chain Reaction (PCR)
Particle Image Velocimetry (PIV)
Infrared Spectroscopy (IR)
SDGs
Type
thesis
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