Journal article
Towards a portable microchip system with integrated thermal control and polymer waveguides for real-time PCR
Department of Micro- and Nanotechnology, Technical University of Denmark1
ChemLabChip Group, LabChip Section, Department of Micro- and Nanotechnology, Technical University of Denmark2
LabChip Section, Department of Micro- and Nanotechnology, Technical University of Denmark3
Section of Poultry Diseases, Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark4
Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark5
National Veterinary Institute, Technical University of Denmark6
BioLabChip Group, LabChip Section, Department of Micro- and Nanotechnology, Technical University of Denmark7
A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. The integrated polymer optical system for real-time monitoring of PCR was fabricated in the same SU-8 layer as the PCR chamber, without additional masking steps. Two suitable DNA binding dyes, SYTOX Orange and TO-PRO-3, were selected and tested for the real-time PCR processes.
As a model, cadF gene of Campylobacter jejuni has been amplified on the microchip. Using the integrated optical system of the real-time PCR microchip, the measured cycle threshold values of the real-time PCR performed with a dilution series of C. jejuni DNA template (2 to 200 pg/mu l) could be quantitatively detected and compared with a conventional post-PCR analysis (DNA gel electrophoresis).
The presented approach provided reliable real-time quantitative information of the PCR amplification of the targeted gene. With the integrated optical system, the reaction dynamics at any location inside the micro reaction chamber can easily be monitored.
Language: | English |
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Publisher: | WILEY-VCH Verlag |
Year: | 2006 |
Pages: | 5051-5058 |
ISSN: | 15222683 and 01730835 |
Types: | Journal article |
DOI: | 10.1002/elps.200600355 |
ORCIDs: | Bang, Dang Duong and Wolff, Anders |