Ultra fast miniaturized real-time PCR: 40 cycles in less than six minutes
Pavel Neuzil*, Chunyan Zhang, Juergen Pipper, Sharon Oh and Lang Zhuo
Institute of Bioengineering and Nanotechnology 31 Biopolis Way, The Nanos, #04-01, Singapore 138669
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°C s1 and a cooling rate of 125°C s1. 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.
Nucleic Acids Research 2006 34(11):e77; doi:10.1093/nar/gkl416