Rapid DNA Amplification in Bouyancy Driven Closed Loop Microfluidic Systems

摘要

Bouyancy driven polymerase chain reaction (PCR) is emerging as a simplified version of thermally driven bio-analysis systems. In this paper, we have designed, fabricated and modeled a simplified closed loop thermocycler (CLTC) that can perform PCR in as little as 10 mins. Robustness and generality of the CLTC was demonstrated by performing PCR amplification of single as well as multiplex systems containing 5 different human respiratory infection associated virus targets. We present a quantitative analysis of PCR yields and observe consistent performance over several orders of magnitude of initial template loading dilution. We also demonstrate the use of convective flows in a variety of closed loop systems designed to create extensional flow fields and to execute tunable thermocycling and pumping operations in a format suitable for integration into miniaturized chemical and biochemical analysis systems.