DNA amplification using the polymerase chain reaction (PCR) is an important tool in biotechnology, pathogen surveillance in food, medical and forensic science etc. The PCR technique is now an important part of the research in and development of miniaturized biochemical analysis systems. However, reduced or no DNA amplification at all is an important challenge for microfabricated PCR devices due to a negative interaction between PCR chemicals and the surrounding environment, i.e. the materials encapsulating the PCR mix. Materials of special interest regarding PCR compatibility are silicon, glass and polymers, which are important in the fabrication of microelectromechanical systems (MEMS), micro total analysis systems (µTAS) and lab-on-a-chip (LOC) systems. The PCR inhibition effect is a particularly important phenomenon in microsystems due to an increased surface-to-volume ratio which enhances the possibility of interaction between the surfaces and ingredients in the PCR mixture. By proper surface treatment the PCR reaction can be facilitated and in this paper we present a systematic and quantitative study of the impact on the PCR compatibility of a chemical and a biological surface treatment. The chemical treatments are based on the silanizing agent dichlordimethylsilane [(CH 3 ) 2 SiCl 2 ]], while the biological treatment is based on the protein bovine serum albumin (BSA). We present a simple model system for the investigation of the PCR compatibility of three widely used materials in microfabrication, namely silicon, glass and SU-8. The impact on PCR performance, measured by means of PCR efficiency, of untreated as well as chemically and biologically treated materials is studied. We show a convenient method of assessing the PCR compatibility of silicon, glass and SU-8 with a degree of information not presented before.
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.
A novel real-time PCR microchip platform with integrated thermal system and polymer waveguides has been developed. By using the integrated optical system of the real-time PCR chip, cadF -a virulence gene of Campylobacter jejuni, could specifically be detected. Two different DNA binding dyes, SYTOX Orange and TO-PRO-3, were added to the PCR mixture to realize the real-time PCR. The presented approach shows reliable real-time quantitative information of the PCR amplification of the targeted gene.
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