Abstract
Abstract Flow-through PCR devices for performing rapid and small-volume DNA amplification on a single chip have attracted great interest. Flow-through DNA amplification was performed by moving PCR solution as a liquid plug through three individual temperature zones. Since precise control of the temperature setting is the most important technique for successful DNA amplification, real-time temperature analyses of the moving fluid were investigated using a polyolefin pressure-sensitive adhesive (PPSA) film to cover the microchannel. The temperature profile at 20 °C during annealing phase was the closest to the recommended conditions for PCR compared to the profile from 55 to 20 °C, and it showed the highest amplification of the tested regimes. The microchannel design was optimized using an infrared (IR) thermal imager to significantly increase the fluorescence intensity of the amplified products. A flow time of five to six seconds per cycle resulted in a temperature profile close to the recommended thermal gradient. These studies resulted in effective findings for simple and rapid amplification by moving a PCR solution as liquid-plug on a single chip.
Published Version
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