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A novel real-time PCR machine with a miniature spectrometer for fluorescence sensing in a micro liter volume glass capillary

Identifieur interne : 000083 ( PascalFrancis/Curation ); précédent : 000082; suivant : 000084

A novel real-time PCR machine with a miniature spectrometer for fluorescence sensing in a micro liter volume glass capillary

Auteurs : D. S. Lee [Taïwan] ; M. H. Wu ; U. Ramesh ; C. W. Lin ; T. M. Lee ; P. H. Chen

Source :

RBID : Pascal:04-0244080

Descripteurs français

English descriptors

Abstract

This study sets up a prototype of real-time polymerase chain reaction (RT-PCR) machine that employs a miniature spectrometer for detecting the emission of fluorescence intensity from RT-PCR mix in a micro liter volume glass capillary. The RT-PCR machine is one of the major instruments for SARS virus test during the outbreak in Asia in early 2003. Comparing with traditional RT-PCR machine with discrete channels fluorescence wavelength detection, the prototype can provide continuous wavelength detection and can be employed for multiplex DNA quantification. However, only one HBV SC 11 DNA template with the SYBR Green I labeling dye were used in this study to compare DNA quantification accuracy and reproducibility of the present system and the commercial system. The two machines have different optical engine arrangement and so two separate analytical models were proposed to predict the fluorescence intensity from the RT-PCR mix during thermal cycling for the machines. Predicted results agree well with the measured data for both machines. From the predicted results, different approaches should be adopted to determine the initial DNA concentration for quantification from time recorded history of the fluorescence intensity. Measured results illustrate that the RT-PCR prototype has the same accuracy for DNA quantification and reproducibility within five intra-assay samples as compared with the commercial machine. © 2004 Elsevier B.V. All rights reserved.
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A08 01  1  ENG  @1 A novel real-time PCR machine with a miniature spectrometer for fluorescence sensing in a micro liter volume glass capillary
A11 01  1    @1 LEE (D. S.)
A11 02  1    @1 WU (M. H.)
A11 03  1    @1 RAMESH (U.)
A11 04  1    @1 LIN (C. W.)
A11 05  1    @1 LEE (T. M.)
A11 06  1    @1 CHEN (P. H.)
A14 01      @1 Mechanical Engineering Department National Taiwan University @2 Taipei 10617 @3 TWN @Z 1 aut.
A20       @1 401-410
A21       @1 2004
A23 01      @0 ENG
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C01 01    ENG  @0 This study sets up a prototype of real-time polymerase chain reaction (RT-PCR) machine that employs a miniature spectrometer for detecting the emission of fluorescence intensity from RT-PCR mix in a micro liter volume glass capillary. The RT-PCR machine is one of the major instruments for SARS virus test during the outbreak in Asia in early 2003. Comparing with traditional RT-PCR machine with discrete channels fluorescence wavelength detection, the prototype can provide continuous wavelength detection and can be employed for multiplex DNA quantification. However, only one HBV SC 11 DNA template with the SYBR Green I labeling dye were used in this study to compare DNA quantification accuracy and reproducibility of the present system and the commercial system. The two machines have different optical engine arrangement and so two separate analytical models were proposed to predict the fluorescence intensity from the RT-PCR mix during thermal cycling for the machines. Predicted results agree well with the measured data for both machines. From the predicted results, different approaches should be adopted to determine the initial DNA concentration for quantification from time recorded history of the fluorescence intensity. Measured results illustrate that the RT-PCR prototype has the same accuracy for DNA quantification and reproducibility within five intra-assay samples as compared with the commercial machine. © 2004 Elsevier B.V. All rights reserved.
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C03 01  1  ENG  @0 RT-PCR @4 INC
C03 02  1  ENG  @0 Real-time polymerase chain reaction @4 INC
C03 03  1  ENG  @0 Discrete channel fluorescence detection @4 INC
C03 04  1  ENG  @0 Continuous wavelength detection @4 INC
C03 05  1  FRE  @0 Théorie
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C03 06  1  FRE  @0 Enzyme
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C03 08  1  FRE  @0 DNA
C03 08  1  ENG  @0 DNA
C03 09  1  FRE  @0 Fluorescence
C03 09  1  ENG  @0 Fluorescence
C03 10  1  FRE  @0 Electrophorèse
C03 10  1  ENG  @0 Electrophoresis
C03 11  1  FRE  @0 Photodiode
C03 11  1  ENG  @0 Photodiodes
C03 12  1  FRE  @0 Sang
C03 12  1  ENG  @0 Blood
C03 13  1  FRE  @0 Colorant
C03 13  1  ENG  @0 Dyes
C03 14  1  FRE  @0 Spectromètre
C03 14  1  ENG  @0 Spectrometers
C03 15  1  FRE  @0 Capteur @3 P
C03 15  1  ENG  @0 Sensors @3 P
N21       @1 159

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Pascal:04-0244080

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