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

Abstract

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 µL. 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.

Keywords:

• DNA amplification
• Miniaturisation
• Silicon micromachining

Acknowledgements

This work was funded by Enterprise Ireland Research Innovation Fund Project Grant EI/RIF/2001/071. We acknowledge the assistance of Ms Marion MacSweeney for wire bonding devices.