Development of a Novel Multiplexing Real-Time PCR Machine for DNA Quantification
Date Issued
2004
Date
2004
Author(s)
Wu, Meng-Hsun
DOI
zh-TW
Abstract
This study sets up a prototype of Multiplexing Real-Time polymerase chain reaction (PCR) machine that employs a xenon light source for exciting fluorescence and a miniature spectrometer for detecting the emission fluorescence intensity from Real-Time PCR mix in a ml volume glass capillary. Compared with the traditional Real-Time PCR machine with discrete channels fluorescence wavelength detection, the prototype can provide continuous wavelength detection and can be employed for multiplex DNA quantification. To compare the DNA quantification accuracy and reproducibility of the present system with those of the commercial system, we use a HBV DNA template with LC-Red 640 dye and an Internal Control template with LC-Red 705 dye in this study. The results of this experiment reveal that the Real-Time PCR prototype yields more amplification quantity than the commercial system. As the initial copies of template DNA increase every ten times, Cp decreases by a value of 3.9. Therefore, the number of template DNA increases by 1.8 times each thermal cycle. The results also reveal that this prototype has the same accuracy for DNA quantification and the reproducibility within five intra assay samples as the commercial system.
Subjects
全波段氙氣燈
連續式波段螢光檢測系統
連鎖反應
複式即時定量聚合酶
spectrometer
xenon light source
continuous wavelength fluorescence detection chann
Multiplexing Real-Time polymerase chain reaction (
Type
thesis
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