Hofmann, MH,Akkoyun, A,Flynn, R,Mathewson, A,Berney, H,Sheehan, MM

2004

September

International Journal of Environmental Analytical Chemistry

Development of pcr conditions in a silicon microreactor DNA-amplification device

Validated

()

DNA amplification miniaturisation silicon micromachining POLYMERASE-CHAIN-REACTION THERMAL CYCLER CHIP

84

821

833

A silicon microsystem was developed which functions as a miniaturised DNA-amplification device. The system represents a technology platform for performing a polymerase chain reaction (PCR) with reduced volumes of 7 muL. The silicon microreactor was fabricated using silicon bulk micromachining, and a platinum heater was fabricated on a Pyrex substrate. A miniaturised DNA-amplification system permitted rapid heating and cooling, and shorter reaction times of 30 min were achieved. In this work, biocompatibility issues are addressed; conditions for efficient PCR in a silicon-based microreactor are established for the amplification of 500 bp DNA from the Escherichia coli bacteriophage Lambda; and the conditions are verified by amplifying a 255 bp region from the Mycobacterium tuberculosis rpoB gene. This work describes the PCR volume scale down experiments that were conducted and concentrations of the reactants; Taq polymerase, oligonucleotides, MgCl2 and template DNA were determined for DNA-amplification reactions with this novel device.

10.1080/0306731042000268143