Abstract

Multiple microfluidic devices incorporating real-time polymerase chain reaction (PCR) for the quantification of DNA targets have been reported. They show features such as fast analysis time, reduced reagent consumption, and portability. Integration of real-time PCR in a centrifugal microfluidic disc has been also proposed due to the ease of automation and parallelization of microfluidic operations. Previously, it has been demonstrated the implementation of thermal convective PCR (TC-PCR) in a CD using a ring-structured microchannel. In this case, the microchannel is supplied with the temperature necessary to perform the complete thermal cycle: Denaturation, annealing, and extension. The flow generated by thermal convection, in combination with the centrifugal force moves the sample through the entire microchannel due to the difference in densities. In this work, we present the implementation in a CD of an electrochemical sensor combined with TC-PCR to quantify the amplification yield in real-time. We present advances related to the calibration of the electrochemical sensor, and the implementation of the TC-PCR in a CD as well as a preliminary simulation in COMSOL.

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