Design, Fabrication, and Testing of a Continuous Flow, Submicroliter Scale PCR for the Analysis of Plant Genomes

Abstract

Abstract In recent years, many micromachined polymerase chain reaction (PCR) chips have been reported. These miniaturized PCR chips have great advantages such as the significant reduction in reagent costs and vastly reduced reaction times over the conventional PCR reactions. In this paper a micro analysis system that will allow submicro-liter scale, continuous-flow PCR to be conducted in a glass chip has been presented. This glass chip is fabricated through thermally bonding two pyrex 7740 glass wafers. One pyrex wafer is etched to form the 20-cycle microchannel with 80 microns wide and 30 microns deep. Another pyrex wafer is thermally bonded to the etched wafer to produce closed a continuous microchannel for PCR. The total length of the microchannel is 0.5 m. This PCR chip has a significant reagent reduction with a volume of less than 1 micro-liter. With 1 micro-liter reagent we obtain a total reaction time of 0.5 min to 3 min depending on various flow rates. This analytical chip is fabricated using standard micromachining techniques with a total size of 56 mm × 24 mm × 1 mm. The advantages of this chip include very small quantities of reagent needed, rapid thermal cycling, standard micromachining processes and batch fabrication which result in a significant cost reduction.