Abstract: A new continuous-flow PCR microfluidics was successfully designed and constructed, in which several factors that might affect the continuous-flow PCR amplification had been investigated in detail, including annealing temperature, concentrations of some PCR reagents (forward/reverse primers, Mg2+, dNTPs and DNA polymerase enzyme) and flow rates of PCR solution in the capillary microchannel. It has been found that the ways these reagents affect the continuous-flow PCR are different from those that affect the conventional PCR, and that these reagents within a wider concentration range won't result in the non-specific PCR amplification. In addition, the 249 bp β-actin gene fragments could be successfully amplified on the present PCR microfluidics within 15 min, which was much faster than the reaction time with the conventional PCR machine. By using the flexible thin film heaters with low thermal mass to maintain the three thermostable zones, the energy consumption of a 33 continuous-flow PCR cycles is less than 0.0088 kWh, which may make the present continuous-flow PCR microfluidics portable.