A temperature control method for shortening thermal cycling time to achieve rapid polymerase chain reaction (PCR) in a disposable polymer microfluidic device

Minqiang Bu, Ivan R. Perch-Nielsen, Karen Skotte Sørensen, Julia Skov, Sun Yi, Dang Duong Bang, Michael E Pedersen, Mikkel Fougt Hansen, Anders Wolff

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Abstract

We present a temperature control method capable of effectively shortening the thermal cycling time of polymerase chain reaction (PCR) in a disposable polymer microfluidic device with an external heater and a temperature sensor. The method employs optimized temperature overshooting and undershooting steps to achieve a rapid ramping between the temperature steps for DNA denaturation, annealing and extension. The temperature dynamics within the microfluidic PCR chamber was characterized and the overshooting and undershooting parameters were optimized using the temperature-dependent fluorescence signal from Rhodamine B. The method was validated with the PCR amplification of mecA gene (162 bp) from methicillin-resistant Staphylococcus aureus bacterium (MRSA), where the time for 30 cycles was reduced from 50 min (without over- and undershooting) to 20 min.
Original languageEnglish
Article number074002
JournalJournal of Micromechanics and Microengineering
Volume23
Issue number7
Number of pages9
ISSN0960-1317
DOIs
Publication statusPublished - 2013

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