Towards a portable microchip system with integrated thermal control and polymer waveguides for real-time PCR.
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/microL) 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.
Wang
Zhenyu
Z
MIC - Department of Micro and Nanotechnology, Technical University of Denmark, Lyngby, Denmark.
Sekulovic
Andrea
A
Kutter
Jörg P
JP
Bang
Dang D
DD
Wolff
Anders
A
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification; D002232:Carbocyanines / Q000032:analysis; D002352:Carrier Proteins / Q000235:genetics; D004247:DNA / Q000032:analysis* / Q000096:biosynthesis; D004269:DNA, Bacterial / Q000032:analysis / Q000096:biosynthesis; D046209:Electrophoresis, Microchip / Q000295:instrumentation*; D008904:Miniaturization; D055095:Optics and Photonics; D009930:Organic Chemicals / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000295:instrumentation* / Q000379:methods*; D011108:Polymers / Q000737:chemistry
D001425:Bacterial Outer Membrane Proteins; C107872:CadF protein, Campylobacter jejuni; D002232:Carbocyanines; D002352:Carrier Proteins; D004269:DNA, Bacterial; D009930:Organic Chemicals; D011108:Polymers; C416802:SYTOX Orange dye; C098830:TO-PRO-3; D004247:DNA