Neuzil Pavel, Zhang Chunyan, Pipper Juergen, Oh Sharon, Zhuo Lang
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, The Nanos, #04-01, Singapore 138669.
Nucleic Acids Res. 2006 Jun 28;34(11):e77. doi: 10.1093/nar/gkl416.
We have designed, fabricated and tested a real-time PCR chip capable of conducting one thermal cycle in 8.5 s. This corresponds to 40 cycles of PCR in 5 min and 40 s. The PCR system was made of silicon micromachined into the shape of a cantilever terminated with a disc. The thin film heater and a temperature sensor were placed on the disc perimeter. Due to the system's thermal constant of 0.27 s, we have achieved a heating rate of 175 degrees C s(-1) and a cooling rate of -125 degrees C s(-1). A PCR sample encapsulated with mineral oil was dispensed onto a glass cover slip placed on the silicon disc. The PCR cycle time was then determined by heat transfer through the glass, which took only 0.5 s. A real-time PCR sample with a volume of 100 nl was tested using a FAM probe. As the single PCR device occupied an area of only a few square millimeters, devices could be combined into a parallel system to increase throughput.
我们设计、制造并测试了一种能够在8.5秒内完成一个热循环的实时PCR芯片。这相当于在5分40秒内完成40个PCR循环。该PCR系统由硅微机械加工成悬臂形状，并在末端带有一个圆盘。薄膜加热器和温度传感器放置在圆盘周边。由于该系统的热常数为0.27秒，我们实现了175℃/秒的加热速率和-125℃/秒的冷却速率。将封装有矿物油的PCR样品滴加到放置在硅盘上的玻璃盖玻片上。然后，PCR循环时间由通过玻璃的热传递决定，这仅需0.5秒。使用FAM探针测试了体积为100 nl的实时PCR样品。由于单个PCR装置仅占据几平方毫米的面积，因此可以将多个装置组合成一个并行系统以提高通量。
