A High Throughput Microfluidic Thermal Reactor Available to Purchase

A high throughput microfluidic system including a 96 continuous flow (CF) thermal reactors and a multi-zone thermal control system was designed and fabricated. An infrared camera (IR) was used to analyze and verify the uniformity of the temperature distribution. Temperature variations from the nominal values were ±2°C in the denaturation zone and ±1°C in the renaturation and extension zones. Six different DNA fragments, with lengths ranging from 99 bp to 997 bp, were obtained from a λ-DNA template, each with a distinct renaturation temperature. As an initial demonstration of the biochemical performance of the polymerase chain reaction (PCR) reactor arrays, a column device comprised of eight 25-cycle CFPCRs was used to amplify identical PCR cocktails for each DNA fragment simultaneously at a flow velocity of 2 mm/s. All but the 997 bp were successfully amplified. Yields for the 99 bp fragment varied from 15%–36% of the amplicon on block thermal cycler. In a second experiment, a row device composed of six 25-cycle CFPCRs was used to successfully, simultaneously amplify cocktails for all six DNA fragment lengths at a flow velocity of 1 mm/s in parallel. Each device in the row had a distinct renaturation temperature to match the DNA fragment length. The parallel thermal arrays can be used in modular systems including both PCR for amplification and, for example, ligase detection reaction (LDR) for mutation detection.