A thermostat chip of indium tin oxide glass substrate for static polymerase chain reaction and in situ real time fluorescence monitoring |
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Authors: | Wu Zhi-Yong Chen Kun Qu Bai-Yan Tian Xiao-Xi Wang Xiao-Jie Fang Fang |
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Institution: | Research Center of Analytical Science, Northeastern University, Wenhua Road 3-11, Shenyang 110004, PR China |
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Abstract: | A thermostat chip of indium-tin oxide glass substrate for static chip polymerase chain reaction (PCR) is, for the first time, introduced in this paper. The transparent conductive layer was used as an electro-heating element. Pulse width modulation and fuzzy proportional integration-differentiation algorithm were adopted in the temperature programming of the chip. The temperature distribution was investigated, and a dynamic control precision within ±2 °C was achieved. The highest ramping rates were 37 °C s−1 for heating and 8 °C s−1 for cooling with an electric fan. The PCR reaction vials were constructed with polyethylene tubes or poly(dimethylsiloxane) directly on the thermostat chip; the chip had a typical size of 25 mm × 25 mm and a thickness of 1.1 mm. Static chip PCR was successfully demonstrated either in a single vial or in an up to 8-parallel array vials. In situ real time fluorescence monitoring during PCR of a λ DNA fragments (236 bp) with SYBR Green I was demonstrated using a blue light emission diode as a light source and a photomultiplier as a detector. The method proposed here is characterized by open access, easy fabrication and low cost. This work could be the basis for developing a portable real time PCR system with disposable chips for point of care tests. |
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Keywords: | Indium-tin oxide glass Chip thermostat Static chip polymerase chain reaction In situ real time fluorescence monitoring Light emission diode |
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