Real time PCR on disposable PDMS chip with a miniaturized thermal cycler.
This paper presents the design and implementation of a low-cost miniature PCR device consisting of a disposable reactor chip and a miniature thermal cycler. The simple fabrication of the PCR chip by PDMS (Polydimethylsiloxane) does not need micro-machining or photolithography processes. The thermal cycler was built with a thin film heater for heating and a fan for rapid cooling. This device can perform PCR tests in a single well chip or a multiple-well chip. It can run PCR reactions of different volumes to meet specific application requirements. The smallest reaction volume tested in this work is 0.9 microL. In addition, this device fits any standard fluorescence microscope for real time detection, which makes real time PCR affordable for most research labs and clinics with a fluorescence microscope. Real-time PCR of E. coli stx1 has been demonstrated with the device described.
Xiang
Q
Q
Department of Mechanical and Industrial Engineering, University of Toronto, 5 King's College Road, Toronto M5S 3G8, Canada.
Xu
B
B
Fu
R
R
Li
D
D
D003199:Computer Systems; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D004129:Dimethylpolysiloxanes; D004209:Disposable Equipment; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006361:Heating / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008904:Miniaturization; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D012828:Silicones; D013696:Temperature
D004269:DNA, Bacterial; D004129:Dimethylpolysiloxanes; D012828:Silicones; C013830:baysilon