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

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...

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Bibliographic Details
Published in:Journal of micromechanics and microengineering 2013-07, Vol.23 (7), p.74002-9
Main Authors: Bu, Minqiang, Perch-Nielsen, Ivan R, Sørensen, Karen S, Skov, Julia, Sun, Yi, Duong Bang, Dang, Pedersen, Michael E, Hansen, Mikkel F, Wolff, Anders
Format: Article
Language:English
Subjects:
Annealing
Biological and medical sciences
Devices
Exact sciences and technology
General equipment and techniques
Heaters
Instruments, apparatus, components and techniques common to several branches of physics and astronomy
Medical sciences
Microfluidics
Physics
Polymer microfluidic device
Polymerase chain reaction
Polymerase chain reaction (PCR)
Radiotherapy. Instrumental treatment. Physiotherapy. Reeducation. Rehabilitation, orthophony, crenotherapy. Diet therapy and various other treatments (general aspects)
Sensors (chemical, optical, electrical, movement, gas, etc.)
remote sensing
Staphylococcus aureus
Technology. Biomaterials. Equipments. Material. Instrumentation
Temperature control
Temperature control method
Thermal cycling
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Summary: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.
ISSN:0960-1317
1361-6439
DOI:10.1088/0960-1317/23/7/074002