Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter VolumesReport as inadecuate




Chip-Oriented Fluorimeter Design and Detection System Development for DNA Quantification in Nano-Liter Volumes - Download this document for free, or read online. Document in PDF available to download.

Department of Energy and Refrigerating Air-Conditioning Engineering, National Taipei University of Technology, Taipei 10608, Taiwan





*

Author to whom correspondence should be addressed.



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. View Full-Text

Keywords: polymerase chain reaction PCR; DNA quantification; chip-oriented fluorimeter design; real-time PCR on a chip system; S-N ratio polymerase chain reaction PCR; DNA quantification; chip-oriented fluorimeter design; real-time PCR on a chip system; S-N ratio





Author: Da-Sheng Lee * and Ming-Hui Chen

Source: http://mdpi.com/



DOWNLOAD PDF




Related documents