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Towards a portable microchip system with integrated thermal control and polymer waveguides for real‐time PCR
Author(s) -
Wang Zhenyu,
Sekulovic Andrea,
Kutter Jörg P.,
Bang Dang D.,
Wolff Anders
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200600355
Subject(s) - campylobacter jejuni , microfluidics , polymer , electrophoresis , real time polymerase chain reaction , materials science , chromatography , analytical chemistry (journal) , chemistry , nanotechnology , gene , biology , genetics , biochemistry , bacteria , composite material
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, cad F 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/µL) 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.