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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WO
WIPO (PCT)
Prior art keywords
amplificates
cytosine methylation
single strands
detecting cytosine
strands
Prior art date
Application number
PCT/DE2002/002433
Other languages
English (en)
French (fr)
Other versions
WO2003002760A2 (de
Inventor
Juergen Distler
Erik Leu
Original Assignee
Epigenomics Ag
Juergen Distler
Erik Leu
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 Epigenomics Ag, Juergen Distler, Erik Leu filed Critical Epigenomics Ag
Priority to US10/482,433 priority Critical patent/US20040265814A1/en
Priority to EP02754310A priority patent/EP1404879A2/de
Publication of WO2003002760A2 publication Critical patent/WO2003002760A2/de
Publication of WO2003002760A3 publication Critical patent/WO2003002760A3/de

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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

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 WO2003002760A2 (de)

Priority Applications (2)

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

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)

Application Number Title Priority Date Filing Date
PCT/DE2002/002433 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 (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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

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US8076063B2 (en) 2000-02-07 2011-12-13 Illumina, Inc. Multiplexed methylation detection methods
US7955794B2 (en) 2000-09-21 2011-06-07 Illumina, Inc. Multiplex nucleic acid reactions
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
ATE462017T1 (de) * 2002-12-02 2010-04-15 Illumina Cambridge Ltd Bestimmung der methylierung von nukleinsäuresequenzen
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
WO2010048337A2 (en) 2008-10-22 2010-04-29 Illumina, Inc. Preservation of information related to genomic dna methylation
WO2011041695A1 (en) * 2009-10-02 2011-04-07 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
CN111057752A (zh) * 2014-02-28 2020-04-24 国立研究开发法人国立癌研究中心 离子交换色谱的应用和pcr引物的应用

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9394565B2 (en) 2003-09-05 2016-07-19 Agena Bioscience, Inc. Allele-specific sequence variation analysis
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
DE10132212B4 (de) 2005-11-24
DE10132212A1 (de) 2003-03-06
WO2003002760A2 (de) 2003-01-09
EP1404879A2 (de) 2004-04-07
US20040265814A1 (en) 2004-12-30

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