Disposable real-time microPCR device: lab-on-a-chip at a low cost.
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.
Neuzil
Pavel
P
Institute of Bioengineering and Nanotechnology, 31 Biopolis Way, Nanos #04-01, Singapore 138669. pneuzil@ibn.a-star.edu.sg
Pipper
Juergen
J
Hsieh
Tseng Ming
TM
D003365:Costs and Cost Analysis; D004247:DNA / Q000032:analysis; D019075:Electrophoresis, Capillary; D004867:Equipment Design / Q000295:instrumentation*; D007753:Laboratories / Q000191:economics*; D008832:Microchemistry / Q000295:instrumentation* / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D013696:Temperature; D013997:Time Factors
D004247:DNA