Continuous flow, submicro-liter scale PCR chip for DNA amplification

摘要

Polymerase chain reaction (PCR) is a well-described method for selective identical replication of DNA molecules. in recent years, many micromachined PCR chips have been reported. These miniaturized PCR chips have great advantages such as a significant reduction in reagent costs and vastly reduced reaction time over the conventional PCR devices. 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 achieved through thermally bonding two pyrex 7740 glass wafers. One pyrex wafer is etched to form a 20-cycle microchannel of 80 micron wide and 30 micron deep. The other pyrex wafer with microheaters is thermally bonded to the microchannel wafer to produce a closed continuous microchannel for PCR. The total length of the microchannel is 0.5 m. The size of this device is 56 mm * 24 mm * 1 mm. Three reaction temperatures are controlled by three PID controllers. This PCR chip has a significant reagent reduction with a volume of less than 1 micro-liter. With 1 micro-liter reagent, we get total reaction time of 0.5 min to 3 min depending on various flow rates. This analysis chip is fabricated using standard micromachining techniques. The advantages of this chip include small quantities of reagent needed, high throughput, rapid thermal cycling, and batch micro-fabrication resulting in a significant cost reduction.