Microfabricated device for DNA and RNA amplification by continuous-flow polymerase chain reaction and reverse transcription-polymerase chain reaction with cycle number selection.

摘要: We have developed a high-throughput microfabricated, reusable glass chip for the functional integration of reverse transcription (RT) and polymerase chain reaction (PCR) in continuous-flow mode. The allows selection number amplification cycles. A single microchannel network was etched that defines four distinct zones, one RT three PCR (denaturation, annealing, extension). zone temperatures were controlled by placing over heating blocks. Samples reagents pumped continuously through appropriate access holes. Outlet channels after cycles 20, 25, 30, 35, 40 product collection. surface-to-volume ratio channel is 57 mm-1 depth 55 μm, both which allow very rapid heat transfer. As result, we able to collect 30 only 6 min. Products collected 0.2-mL tubes analyzed agarose gel electrophoresis ethidium bromide staining....