Chip-oriented fluorimeter design and detection system development for DNA quantification in nano-liter volumes.
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 10(3) to 10(5) copies per reaction.
Department of Energy and Refrigerating Air-Conditioning Engineering, National Taipei University of Technology, Taipei 10608, Taiwan. f11167@ntut.edu.tw
Da-Sheng
DS
Lee
Ming-Hui
MH
Chen
D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005470:Fluorometry / Q000295:instrumentation*; D046228:Microarray Analysis / Q000295:instrumentation*; D008832:Microchemistry / Q000295:instrumentation*; D036103:Nanotechnology / Q000295:instrumentation; D016133:Polymerase Chain Reaction / Q000295:instrumentation*; D014159:Transducers
D004247:DNA