Continuous-flow polymerase chain reaction microfluidics based on polytetrafluoethylene capillary

Abstract

The polytetrafluoroethylene (PTFE) capillary-based continuous-flow polymerase chain reaction (PCR) microfluidics were constructed by the flexible thin film heaters to form three constant temperature zones necessary for a PCR amplification, in which the PCR mixture could flow through the capillary mcirochannel continuously, thus leading to the successful completion of PCR exponential amplification. The effects of various concentrations of bovine serum albumin (BSA) in a 25-μl PCR mixture and flow rates of PCR mixture in the capillary microchannel, on continuous-flow PCR amplification, had been discussed in detail, where a PCR amplification of 249-bp human β-actin gene fragments was successfully performed within 15 and even 10 minutes, which is several times faster than the reaction time required (1–2 hours) with the conventional PCR machine.