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WO2003002760A3 - Verfahren zum nachweis von cytosin-methylierung durch vergleichende analyse der einzelstränge von amplifikaten - Google Patents

Verfahren zum nachweis von cytosin-methylierung durch vergleichende analyse der einzelstränge von amplifikaten Download PDF

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Publication number
WO2003002760A3
WO2003002760A3 PCT/DE2002/002433 DE0202433W WO03002760A3 WO 2003002760 A3 WO2003002760 A3 WO 2003002760A3 DE 0202433 W DE0202433 W DE 0202433W WO 03002760 A3 WO03002760 A3 WO 03002760A3
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WIPO (PCT)
Prior art keywords
amplificates
cytosine methylation
single strands
detecting cytosine
strands
Prior art date
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Ceased
Application number
PCT/DE2002/002433
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English (en)
French (fr)
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WO2003002760A2 (de
Inventor
Juergen Distler
Erik Leu
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Epigenomics AG
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Epigenomics AG
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Filing date
Publication date
Application filed by Epigenomics AG filed Critical Epigenomics AG
Priority to EP02754310A priority Critical patent/EP1404879A2/de
Priority to US10/482,433 priority patent/US20040265814A1/en
Publication of WO2003002760A2 publication Critical patent/WO2003002760A2/de
Publication of WO2003002760A3 publication Critical patent/WO2003002760A3/de
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12Q1/6858—Allele-specific amplification

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Beschrieben wird ein Verfahren zum Nachweis von Cytosin-Methylierung in DNA-Proben. Eine genomische DNA Probe wird chemisch, bevorzugt mit einem Bisulfit (=Disulfit, Hydrogensulfit), derart behandelt, dass Cytosin in Uracil umgewandelt wird, während 5-Methylcytosin unverändert bleibt. Abschnitte der Proben-DNA werden mit mindestens 2 Primern in einer Polymerasereaktion, bevorzugt einer Polymerasekettenreaktion, amplifiziert. Zuletzt werden die Fragmente hinsichtlich der Basenzusammensetzung jeweils der beiden komplementären Stränge des Amplifikates untersucht, wobei aus dem Unterschied im Molekulargewicht der beiden Stränge auf den Methylierungsstatus in dem amplifizierten Abschnitt der genomischen DNA-Probe geschlossen wird.
PCT/DE2002/002433 2001-06-27 2002-06-27 Verfahren zum nachweis von cytosin-methylierung durch vergleichende analyse der einzelstränge von amplifikaten Ceased WO2003002760A2 (de)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP02754310A EP1404879A2 (de) 2001-06-27 2002-06-27 Verfahren zum nachweis von cytosin-methylierung durch vergleichende analyse der einzelstränge von amplifikaten
US10/482,433 US20040265814A1 (en) 2001-06-27 2002-06-27 Method for detecting cytosine methylation by comparatively analysing single strands of amplificates

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE10132212A DE10132212B4 (de) 2001-06-27 2001-06-27 Verfahren zum Nachweis von Cytosin-Methylierung durch vergleichende Analyse der Einzelstränge von Amplifikaten
DE10132212.7 2001-06-27

Publications (2)

Publication Number Publication Date
WO2003002760A2 WO2003002760A2 (de) 2003-01-09
WO2003002760A3 true WO2003002760A3 (de) 2003-10-09

Family

ID=7690451

Family Applications (1)

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PCT/DE2002/002433 Ceased WO2003002760A2 (de) 2001-06-27 2002-06-27 Verfahren zum nachweis von cytosin-methylierung durch vergleichende analyse der einzelstränge von amplifikaten

Country Status (4)

Country Link
US (1) US20040265814A1 (de)
EP (1) EP1404879A2 (de)
DE (1) DE10132212B4 (de)
WO (1) WO2003002760A2 (de)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7332275B2 (en) 1999-10-13 2008-02-19 Sequenom, Inc. Methods for detecting methylated nucleotides
US7608394B2 (en) 2004-03-26 2009-10-27 Sequenom, Inc. Methods and compositions for phenotype identification based on nucleic acid methylation
US9249456B2 (en) 2004-03-26 2016-02-02 Agena Bioscience, Inc. Base specific cleavage of methylation-specific amplification products in combination with mass analysis
US9394565B2 (en) 2003-09-05 2016-07-19 Agena Bioscience, Inc. Allele-specific sequence variation analysis

Families Citing this family (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7611869B2 (en) 2000-02-07 2009-11-03 Illumina, Inc. Multiplexed methylation detection methods
US7955794B2 (en) 2000-09-21 2011-06-07 Illumina, Inc. Multiplex nucleic acid reactions
US8076063B2 (en) 2000-02-07 2011-12-13 Illumina, Inc. Multiplexed methylation detection methods
US7582420B2 (en) 2001-07-12 2009-09-01 Illumina, Inc. Multiplex nucleic acid reactions
DE10240746A1 (de) * 2002-09-01 2004-03-18 Epigenomics Ag Verfahren zum Nachweis von Nukleinsäuresequenzen mittels spaltbarer Sondenmoleküle
AU2003290223A1 (en) * 2002-12-02 2004-06-23 Solexa Limited Determination of methylation of nucleic acid sequences
DE10338308B4 (de) * 2003-08-15 2006-10-19 Epigenomics Ag Verfahren zum Nachweis von Cytosin-Methylierungen in DNA
US20060134650A1 (en) * 2004-12-21 2006-06-22 Illumina, Inc. Methylation-sensitive restriction enzyme endonuclease method of whole genome methylation analysis
US7820385B2 (en) 2006-03-22 2010-10-26 The United States Of America As Represented By The Department Of Health And Human Services, Centers For Disease Control And Prevention Method for retaining methylation pattern in globally amplified DNA
WO2008096146A1 (en) 2007-02-07 2008-08-14 Solexa Limited Preparation of templates for methylation analysis
US20080213870A1 (en) * 2007-03-01 2008-09-04 Sean Wuxiong Cao Methods for obtaining modified DNA from a biological specimen
EP2340314B8 (de) 2008-10-22 2015-02-18 Illumina, Inc. Erhaltung von informationen in verbindung mit genomischer dna-methylierung
US20110091882A1 (en) * 2009-10-02 2011-04-21 Ibis Biosciences, Inc. Determination of methylation status of polynucleotides
DE102013009654A1 (de) * 2013-06-07 2014-12-11 Klaus Olek Eine Methode zum Nachweis und zur Unterscheidung von Körperflüssigkeiten aus forensischem Material
EP3112475B1 (de) 2014-02-28 2018-08-15 National Cancer Center Verfahren zur bestimmung der prognose eines nierenzellkarzinoms
CN116626189B (zh) * 2023-05-15 2024-04-30 康龙化成(宁波)科技发展有限公司 一种基于LC-MS对On-DNA化学反应中DNA损伤的分析方法

Citations (2)

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WO1999028498A2 (de) * 1997-11-27 1999-06-10 Epigenomics Gmbh Verfahren zur herstellung komplexer dna-methylierungs-fingerabdrücke
WO2002004686A2 (en) * 2000-07-10 2002-01-17 Epigenx Pharmaceutical, Inc. Detecting methylated cytosine in polynucleotides

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US4060065A (en) * 1973-10-23 1977-11-29 Nissan Motor Company, Limited Exhaust gas recirculation system having means to estimate actual recirculation rate based on intake and exhaust gas temperatures
EP1015746B1 (de) * 1997-04-01 2003-09-10 Robert Bosch Gmbh Einrichtung zum bestimmen der in die zylinder einer brennkraftmaschine mit lader gelangenden luft
US5941927A (en) * 1997-09-17 1999-08-24 Robert Bosch Gmbh Method and apparatus for determining the gas temperature in an internal combustion engine
DE19853398C1 (de) * 1998-11-19 2000-03-16 Epigenomics Gmbh Verfahren zur Identifikation von Cytosin-Methylierungsmustern in genomischer DNA
US6067800A (en) * 1999-01-26 2000-05-30 Ford Global Technologies, Inc. Control method for a variable geometry turbocharger in a diesel engine having exhaust gas recirculation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999028498A2 (de) * 1997-11-27 1999-06-10 Epigenomics Gmbh Verfahren zur herstellung komplexer dna-methylierungs-fingerabdrücke
WO2002004686A2 (en) * 2000-07-10 2002-01-17 Epigenx Pharmaceutical, Inc. Detecting methylated cytosine in polynucleotides

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
FROMMER M ET AL: "A GENOMIC SEQUENCING PROTOCOL THAT YIELDS A POSITIVE DISPLAY OF 5-METHYLCYTOSINE RESIDUES IN INDIVIDUAL DNA STRANDS", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA, NATIONAL ACADEMY OF SCIENCE. WASHINGTON, US, vol. 89, March 1992 (1992-03-01), pages 1827 - 1831, XP002941272, ISSN: 0027-8424 *
OLEK A ET AL: "A modified an improved method for bisulphite based cytosine methylation analysis", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 24, no. 24, 1996, pages 5064 - 5066, XP002106408, ISSN: 0305-1048 *
REIN ET AL: "Identifying 5-methylcytosine and related modifications in DNA genomes", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 26, no. 10, 1998, pages 2255 - 2264, XP002143106, ISSN: 0305-1048 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7332275B2 (en) 1999-10-13 2008-02-19 Sequenom, Inc. Methods for detecting methylated nucleotides
US9394565B2 (en) 2003-09-05 2016-07-19 Agena Bioscience, Inc. Allele-specific sequence variation analysis
US7608394B2 (en) 2004-03-26 2009-10-27 Sequenom, Inc. Methods and compositions for phenotype identification based on nucleic acid methylation
US9249456B2 (en) 2004-03-26 2016-02-02 Agena Bioscience, Inc. Base specific cleavage of methylation-specific amplification products in combination with mass analysis

Also Published As

Publication number Publication date
WO2003002760A2 (de) 2003-01-09
US20040265814A1 (en) 2004-12-30
EP1404879A2 (de) 2004-04-07
DE10132212B4 (de) 2005-11-24
DE10132212A1 (de) 2003-03-06

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