Chip Systems for Analysis of Nucleic Acids with Integrated Amplification and Detection
Analytical methods for the detection of nucleic acids are suitable for a variety of applications as, for example, the analytical applications in that field are diagnosis of infectious or hereditary diseases, analysis of microbial contaminations, determina
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Abstract Analytical methods for the detection of nucleic acids are suitable for a variety of applications as, for example, the analytical applications in that field are diagnosis of infectious or hereditary diseases, analysis of microbial contaminations, determination of family relations, or identification of genetically modified organisms. In many cases, the available number of DNA or RNA molecules is not sufficient for common detection methods based on hybridization processes. The standard procedure for nucleic acid amplification is polymerase chain reaction (PCR). This thermally controlled, enzymatically catalyzed, and cyclically performed process enables the synthesis of small amounts of nucleic acids. In this chapter, a microchip thermocycler with integrated heaters and temperature sensors for droplet-based on-chip PCR is described. The chip is coupled with a
W. Fritzsche (*) • M. Kielpinski • M. Urban • T. Henkel • S. Werres Nanobiophotonics Department, Institute of Photonic Technology (IPHT), PO Box 100 239, 07702 Jena, Germany e-mail: [email protected] R. M€oller Nanobiophotonics Department, Institute of Photonic Technology (IPHT), PO Box 100 239, 07702 Jena, Germany Present address: Analytik Jena AG, Konrad-Zuse-Str. 1, 07745 Jena, Germany S. Wagner Julius-K€uhn-Institute (JKI), Messeweg 11/12, 38104 Braunschweig, Germany M. Riedel Julius-K€uhn-Institute (JKI), Messeweg 11/12, 38104 Braunschweig, Germany Present address: Landesamt f€ ur l€andliche Entwicklung, Landwirtschaft und Flurneuordnung, Steinplatz 1, 15806 Zossen, Germany S. Julich Nanobiophotonics Department, Institute of Photonic Technology (IPHT), PO Box 100 239, 07702 Jena, Germany Present address: microfluidic ChipShop GmbH, Stockholmer Str. 20, 07747 Jena, Germany e-mail: [email protected] W. Fritzsche and J. Popp (eds.), Optical Nano- and Microsystems for Bioanalytics, Springer Series on Chemical Sensors and Biosensors (2012) 10: 289–304 DOI 10.1007/978-3-642-25498-7_11, # Springer-Verlag Berlin Heidelberg 2012
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temperature controlling unit and a device for measuring the fluorescence intensity. Different amounts of initial DNA down to 2 molecules/ml could be successfully detected. The applied template DNA was obtained from different Phytophthora species, a group of phytopathogenic organisms with worldwide significance. Small numbers of cells could also be applied and added directly to the PCR mixture. This was studied successfully with human brain microvascular endothelial (HBME) cells and with spores from Phytophthora. Furthermore, another lab-on-a-chip system is presented that combines on-chip PCR and microarray technology within one chip. The chip consists of a DNA chip with integrated electrode structures and a silicone-based chip with incorporated microchannels and reaction chambers. For temperature management, there are heating elements and temperature sensors connected to the chip. Fluidic transportation is implemented by small
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