WO2004007763A3 - Method and means for quantitative analysis of the number of molecules coding for different genes in cells - Google Patents

Method and means for quantitative analysis of the number of molecules coding for different genes in cells Download PDF

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Publication number
WO2004007763A3
WO2004007763A3 PCT/FR2003/002130 FR0302130W WO2004007763A3 WO 2004007763 A3 WO2004007763 A3 WO 2004007763A3 FR 0302130 W FR0302130 W FR 0302130W WO 2004007763 A3 WO2004007763 A3 WO 2004007763A3
Authority
WO
WIPO (PCT)
Prior art keywords
chain reaction
polymerase chain
genes
cells
primers
Prior art date
Application number
PCT/FR2003/002130
Other languages
French (fr)
Other versions
WO2004007763A2 (en
Inventor
Benedita Rocha
Henrique Veiga-Fernandes
Antonia Peixoto
Original Assignee
Inst Necker
Benedita Rocha
Henrique Veiga-Fernandes
Antonia Peixoto
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inst Necker, Benedita Rocha, Henrique Veiga-Fernandes, Antonia Peixoto filed Critical Inst Necker
Priority to AU2003264700A priority Critical patent/AU2003264700A1/en
Priority to EP03763939A priority patent/EP1552006A2/en
Publication of WO2004007763A2 publication Critical patent/WO2004007763A2/en
Publication of WO2004007763A3 publication Critical patent/WO2004007763A3/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6809—Methods for determination or identification of nucleic acids involving differential detection
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6851—Quantitative amplification

Abstract

The invention concerns a method for in vitro and/or ex vivo quantitative analysis of the number of nucleic acid molecules coding for different genes expressed in cells of living organisms, on a small number of cells, preferably on a single cell, which consists in: performing a reverse transcription and a first polymerase chain reaction (RT-PCR) of mRNA of a small number of lyzed cells, separating the various products of the first polymerase chain reaction to individualize the genes, and performing a second polymerase chain reaction on the resulting separated individual genes. While the first polymerase chain reaction is carried out by means of an association of 5' primers and 3' primers, said 3' primers being identical to the 3' primers used for the reverse transcription, the method consists in using, as reference for calculating the number of copies of genes generated by polymerase chain reaction, the individual cellular DNA coding for the gene concerned, controlling the efficacy of the reverse transcription, by means of a polymerase chain reaction of the cDNA's of one of the genes concerned, in parallel to the same amplification of the mRNA molecules of the same gene, on cellular samples containing the same number of molecules, and evaluating the results of said second polymerase chain reaction.
PCT/FR2003/002130 2002-07-09 2003-07-09 Method and means for quantitative analysis of the number of molecules coding for different genes in cells WO2004007763A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2003264700A AU2003264700A1 (en) 2002-07-09 2003-07-09 Method and means for quantitative analysis of the number of molecules coding for different genes in cells
EP03763939A EP1552006A2 (en) 2002-07-09 2003-07-09 Method and means for quantitative analysis of the number of molecules coding for different genes in cells

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
FR0208593A FR2842211A1 (en) 2002-07-09 2002-07-09 METHOD AND MEANS FOR QUANTITATIVE ANALYSIS OF THE NUMBER OF MOLECULES OF ARNM ENCODING DIFFERENT GENES IN CELLS
FR02/08593 2002-07-09

Publications (2)

Publication Number Publication Date
WO2004007763A2 WO2004007763A2 (en) 2004-01-22
WO2004007763A3 true WO2004007763A3 (en) 2004-06-03

Family

ID=29763659

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/FR2003/002130 WO2004007763A2 (en) 2002-07-09 2003-07-09 Method and means for quantitative analysis of the number of molecules coding for different genes in cells

Country Status (4)

Country Link
EP (1) EP1552006A2 (en)
AU (1) AU2003264700A1 (en)
FR (1) FR2842211A1 (en)
WO (1) WO2004007763A2 (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2874026B1 (en) * 2004-08-04 2006-11-17 Centre Nat Rech Scient Cnrse QUANTIFICATION OF PROTEIN INTEREST MRIAS USING A REAL-TIME RT-PCR REACTION
US8478544B2 (en) 2007-11-21 2013-07-02 Cosmosid Inc. Direct identification and measurement of relative populations of microorganisms with direct DNA sequencing and probabilistic methods
EP2229587B1 (en) * 2007-11-21 2016-08-03 Cosmosid Inc. Genome identification system
EP3972611A4 (en) * 2019-05-21 2023-06-14 Academia Sinica Method for amplifying and detecting ribonucleic acid (rna) fragments

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999054510A2 (en) * 1998-04-23 1999-10-28 Genentech, Inc. Quantitative analysis of gene expression
WO2000020629A2 (en) * 1998-10-05 2000-04-13 Connaught Laboratories Limited Quantitation of rna
EP1138783A2 (en) * 2000-03-31 2001-10-04 Roche Diagnostics GmbH Method for the efficiency-corrected real-time quantification of nucleic acids

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999054510A2 (en) * 1998-04-23 1999-10-28 Genentech, Inc. Quantitative analysis of gene expression
WO2000020629A2 (en) * 1998-10-05 2000-04-13 Connaught Laboratories Limited Quantitation of rna
EP1138783A2 (en) * 2000-03-31 2001-10-04 Roche Diagnostics GmbH Method for the efficiency-corrected real-time quantification of nucleic acids

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
FREEMAN WILLARD M ET AL: "Quantitative RT-PCR: Pitfalls and potential", BIOTECHNIQUES, EATON PUBLISHING, NATICK, US, vol. 26, no. 1, January 1999 (1999-01-01), pages 112 - 118,120-122,124-12, XP002143860, ISSN: 0736-6205 *
See also references of EP1552006A2 *
VEIGA-FERNANDES ET AL.: "Response of naive and memory CD8+ T cells to antigen stimulation in vivo", NATURE IMMUNOLOGY, vol. 1, no. 1, July 2000 (2000-07-01), pages 47 - 53, XP001149835 *
WARTHOE P ET AL.: "Detection and identification of expressed genes by differential display", 1995, COLD SPRING HARNOR LABORATORY PRESS, NY, USA, XP001188140 *

Also Published As

Publication number Publication date
EP1552006A2 (en) 2005-07-13
WO2004007763A2 (en) 2004-01-22
FR2842211A1 (en) 2004-01-16
AU2003264700A1 (en) 2004-02-02

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