Rapid DNA amplification in glass microdevices.
The polymerase chain reaction (PCR) for amplification of DNA has become a very useful tool in scientific research and analytical laboratories, yet conventional techniques are time-consuming, and the reagents are expensive. Miniaturization of this technique has the potential to drastically reduce amplification time and reagent consumption while simultaneously improving the efficiency of the reaction. Increasing the surface area-to-volume ratio using microfluidic reaction chambers allows homogeneous solution temperatures to be achieved much more rapidly than in conventional heating blocks. Employing infrared radiation to selectively heat the reaction solution can additionally reduce the time and energy needed for thermocycling; the reaction container is not heated and can even serve as a heat sink for enhancement of cooling. Microchip systems also provide the potential for fabrication of structures for additional processing steps directly in line with the PCR chamber. Not only can amplification be integrated with product separation and analysis, but sample preparation steps can also be incorporated prior to amplification. The ultimate goal is a miniature total-analysis-system with seamlessly coupled sample-in/answer-out capabilities that consumes very low volumes of reagents and drastically reduces the time for analysis. This chapter will focus on the materials and methods involved in simple straight-channel microchip PCR on glass substrates using non-contact thermocycling.
Easley
Christopher J
CJ
Department of Chemistry, University of Virginia, Charlottesville, VA, USA.
Legendre
Lindsay A
LA
Landers
James P
JP
Ferrance
Jerome P
JP
D001426:Bacterial Proteins / Q000235:genetics; D004247:DNA / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004867:Equipment Design; D005798:Genes, Bacterial; D005898:Glass; D006358:Hot Temperature; D007259:Infrared Rays; D056656:Lab-On-A-Chip Devices; D046208:Microchip Analytical Procedures / Q000379:methods; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods; D011108:Polymers; D012486:Salmonella typhimurium / Q000235:genetics; D013499:Surface Properties
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011108:Polymers; C075518:invA protein, Bacteria; D004247:DNA