Continuous‐flow Polymerase Chain Reaction Microfluidics by Using Spiral Capillary Channel Embedded on Copper

Abstract

This paper presents a novel method for performing polymerase chain reaction (PCR) amplification by using spiral channel fabricated on copper where a transparent polytetrafluoroethylene (PTFE) capillary tube was embedded. The channel with 25 PCR cycles was gradually developed in a spiral manner from inner to outer. The durations of PCR mixture at the denaturation, annealing and extension zones were gradually lengthened at a given flow rate, which may benefit continuous‐flow PCR amplification as the synthesis ability of the Taq polymerase enzyme usually weakens with PCR time. Successful continuous‐flow amplification of DNA fragments has been demonstrated. The PCR products of 249, 500 and 982 bp fragments could be obviously observed when the flow rates of PCR mixture were 7.5, 7.5 and 3.0 mm s⁻¹, respectively, and the required amplification times were about 25, 25, and 62 min, respectively. Besides, the successful segmented‐flow PCR of three samples (249, 500 and 982 bp) has also been reported, which demonstrates the present continuous‐flow PCR microfluidics can be developed for high‐throughput genetic analysis.

Keywords:

• Continuous‐flow
• polymerase chain reaction
• microfluidics
• spiral capillary channel

This work was supported by the National Natural Science Foundation of China (No. 50476088) and the Natural Science Foundation of Guangdong Province (No. 5000729).