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Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter Volumes
Department of Energy and Refrigerating Air-Conditioning Engineering, National Taipei University of Technology, Taipei 10608, Taiwan
* Author to whom correspondence should be addressed.
Received: 23 November 2009; in revised form: 11 December 2009 / Accepted: 22 December 2009 / Published: 28 December 2009
Abstract: The chip-based polymerase chain reaction (PCR) system has been developed in recent years to achieve DNA quantification. Using a microstructure and miniature chip, the volume consumption for a PCR can be reduced to a nano-liter. With high speed cycling and a low reaction volume, the time consumption of one PCR cycle performed on a chip can be reduced. However, most of the presented prototypes employ commercial fluorimeters which are not optimized for fluorescence detection of such a small quantity sample. This limits the performance of DNA quantification, especially low experiment reproducibility. This study discusses the concept of a chip-oriented fluorimeter design. Using the analytical model, the current study analyzes the sensitivity and dynamic range of the fluorimeter to fit the requirements for detecting fluorescence in nano-liter volumes. Through the optimized processes, a real-time PCR on a chip system with only one nano-liter volume test sample is as sensitive as the commercial real-time PCR machine using the sample with twenty micro-liter volumes. The signal to noise (S/N) ratio of a chip system for DNA quantification with hepatitis B virus (HBV) plasmid samples is 3 dB higher. DNA quantification by the miniature chip shows higher reproducibility compared to the commercial machine with respect to samples of initial concentrations from 103 to 105 copies per reaction.
Keywords: polymerase chain reaction (PCR); DNA quantification; chip-oriented fluorimeter design; real-time PCR on a chip system; S/N ratio
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Lee, D.-S.; Chen, M.-H. Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter Volumes. Sensors 2010, 10, 146-166.
Lee D-S, Chen M-H. Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter Volumes. Sensors. 2010; 10(1):146-166.
Lee, Da-Sheng; Chen, Ming-Hui. 2010. "Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter Volumes." Sensors 10, no. 1: 146-166.