Study Of Capillary-Based Continuous-Flow PCR Microfluidics And Dynamical Passivation In Microchannel

Polymerase chain reaction (PCR) is a rapidly developing and widely used nucleic acid amplification technique, which is widely applied onto life science and other related fields. The microfluidics based PCR technology is always one of the hotspots in the increasing development of the microfluidics technology for the biological fields. In the present work, two kinds of new capillary-based PCR microfluidics have been put forward on the basis of the synthetical analysis of the previously reported PCR microfluidics, in which a series of experiments necessary for their biological and chemical performance have been performed.For the continuous-flow PCR microfluidics based on the flexible thin film heaters with low thermal capacity, the performance of temperature controlment and the energy consumption have been evaluated systematically. Besides, a number of factors, which can affect the continuous-flow PCR amplification, have been discussed in detail, including annealing temperature, concentrations of PCR reactants (Mg2+, primers, dNTPs and DNA template, Taq polymerase enzyme), flow rates of PCR solutions through the capillary tube, pretreatment and regeneration of the capillary inner surface and so on. Through these experiments, some important rules for the present continuous-flow PCR microfluidics have been revealed.On the continuous-flow PCR microfluidics by using spiral channel on copper, the experiments were carried out to study the effects of flow rates of PCR solutions in the capillary tube and the initial concentration of human genomic DNA template on continuous-flow PCR amplification. Thus, the performance of thermal cycling rates and PCR amplification limit for the present continuous-flow has been evaluated. In addition, the ability was tested for high-throughput continuous-flow PCR amplification by using the segmented-flow model. For this novel continuous-flow PCR microfluidics, the simplicity in design and the facility in fabrication have been used adequately. However, the size factor of the polytetrafluoroethylene (PTFE)...