A thermostat chip of indium tin oxide glass substrate for static polymerase chain reaction and in situ real time fluorescence monitoring.
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 degrees C was achieved. The highest ramping rates were 37 degrees Cs(-1) for heating and 8 degrees Cs(-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 x 25 mm and a thickness of 1.1mm. 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 lambda DNA fragments (236bp) 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.
Wu
Zhi-Yong
ZY
Research Center of Analytical Science, Northeastern University, Wenhua Road 3-11, Shenyang 110004, PR China. zywu@mail.edu.cn
Chen
Kun
K
Qu
Bai-Yan
BY
Tian
Xiao-Xi
XX
Wang
Xiao-Jie
XJ
Fang
Fang
F
D001483:Base Sequence; D002138:Calibration; D017931:DNA Primers; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013696:Temperature; D017971:Tin Compounds / Q000737:chemistry*
D017931:DNA Primers; D017971:Tin Compounds; C109984:indium tin oxide