Neuzil Pavel, Pipper Juergen, Hsieh Tseng Ming
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Nanos #04-01, Singapore 138669.
Mol Biosyst. 2006 Jun;2(6-7):292-8. doi: 10.1039/b605957k. Epub 2006 May 17.
We have designed, fabricated and tested a real-time micro polymerase chain reaction (microPCR) system. It consists of a microscope glass cover slip placed on top of a micromachined silicon chip integrated with a heater and a temperature sensor. A single microL of a sample containing DNA was placed on the glass and encapsulated with mineral oil to prevent the evaporation of water, thus forming a virtual reaction chamber (VRC). The PCR chip required half a second to heat up from 72 to 94 degrees C and two seconds to cool from 94 to 55 degrees C, corresponding to a cooling rate of -20 K s(-1). The real-time PCR yield was determined by a fluorescence method. The melting curve analysis method as well as capillary electrophoresis was performed to determine the purity of the PCR product. As the glass slip is disposable, cross-contamination from sample to sample is eliminated. The total cost of running the PCR is given by the value of the cover slip and its treatment.
我们设计、制造并测试了一种实时微聚合酶链反应(微PCR)系统。它由放置在与加热器和温度传感器集成的微加工硅芯片顶部的显微镜盖玻片组成。将一微升含有DNA的样品放置在玻璃片上,并用矿物油封装以防止水分蒸发,从而形成一个虚拟反应室(VRC)。PCR芯片从72℃加热到94℃需要半秒,从94℃冷却到55℃需要两秒,冷却速率为-20K s(-1)。实时PCR产量通过荧光法测定。采用熔解曲线分析法以及毛细管电泳法来确定PCR产物的纯度。由于盖玻片是一次性的,消除了样品之间的交叉污染。进行PCR的总成本由盖玻片及其处理的价值给出。