Centrifugo-thermopneumatic fluid control for valving and aliquoting applied to multiplex realtime PCR on off-the-shelf centrifugal thermocycler
15.10.2015
Keller M, Wadle S, Paust N, Dreesen L, Nuese C, Strohmeier O, Zengerle R., von Stetten F
RSC Adv., 2015,5, 89603-89611
We introduce microfluidic automation of geometrically multiplexed real-time PCR to off-the-shelf Rotor-Gene Q thermocyclers (RGQ, QIAGEN GmbH, Hilden, Germany). Compatibility to the very limited flexibility of centrifugal actuation is achieved by using thermal gas compression and expansion for valving and aliquoting. A microfluidic disk segment comprising both new unit operations was used for automation of real-time PCR amplification of Escherichia coli DNA. A comparison with reference reactions in conventional PCR tubes yielded the same PCR efficiency, repeatability, and reproducibility.
