ES2460896T3 - Amplificación de ácido nucleico - Google Patents
Amplificación de ácido nucleico Download PDFInfo
- Publication number
- ES2460896T3 ES2460896T3 ES08800110.2T ES08800110T ES2460896T3 ES 2460896 T3 ES2460896 T3 ES 2460896T3 ES 08800110 T ES08800110 T ES 08800110T ES 2460896 T3 ES2460896 T3 ES 2460896T3
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- primer
- oligonucleotide
- facilitator
- sequence
- facilitator oligonucleotide
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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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
-
- 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
- C12Q1/686—Polymerase chain reaction [PCR]
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (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)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2007905445A AU2007905445A0 (en) | 2007-10-04 | Nucleic acid amplification | |
| AU2007905445 | 2007-10-04 | ||
| PCT/AU2008/001475 WO2009043112A1 (en) | 2007-10-04 | 2008-10-03 | Nucleic acid amplification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ES2460896T3 true ES2460896T3 (es) | 2014-05-14 |
Family
ID=40525777
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES08800110.2T Active ES2460896T3 (es) | 2007-10-04 | 2008-10-03 | Amplificación de ácido nucleico |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US8455197B2 (enExample) |
| EP (1) | EP2207896B1 (enExample) |
| JP (1) | JP5535921B2 (enExample) |
| CN (1) | CN101889096B (enExample) |
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| WO (1) | WO2009043112A1 (enExample) |
Families Citing this family (33)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2271772B1 (en) * | 2008-03-11 | 2014-07-16 | Sequenom, Inc. | Nucleic acid-based tests for prenatal gender determination |
| US8962247B2 (en) * | 2008-09-16 | 2015-02-24 | Sequenom, Inc. | Processes and compositions for methylation-based enrichment of fetal nucleic acid from a maternal sample useful for non invasive prenatal diagnoses |
| US8476013B2 (en) | 2008-09-16 | 2013-07-02 | Sequenom, Inc. | Processes and compositions for methylation-based acid enrichment of fetal nucleic acid from a maternal sample useful for non-invasive prenatal diagnoses |
| EP2454381A1 (en) * | 2009-07-16 | 2012-05-23 | Abbott Laboratories | A method for amplification of nucleic acids |
| EP3088532B1 (en) | 2009-12-22 | 2019-10-30 | Sequenom, Inc. | Processes and kits for identifying aneuploidy |
| WO2012149339A2 (en) | 2011-04-29 | 2012-11-01 | Sequenom, Inc. | Quantification of a minority nucleic acid species |
| CN103930571A (zh) * | 2011-11-17 | 2014-07-16 | 瑞尼克斯有限公司 | 用于选择性分子分析的系统和方法 |
| US9605313B2 (en) | 2012-03-02 | 2017-03-28 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| DK3594366T3 (da) | 2012-05-11 | 2021-09-13 | Clinical Genomics Pty Ltd | Diagnostisk genmarkørpanel |
| US10504613B2 (en) | 2012-12-20 | 2019-12-10 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| US9920361B2 (en) | 2012-05-21 | 2018-03-20 | Sequenom, Inc. | Methods and compositions for analyzing nucleic acid |
| EP2872648B1 (en) | 2012-07-13 | 2019-09-04 | Sequenom, Inc. | Processes and compositions for methylation-based enrichment of fetal nucleic acid from a maternal sample useful for non-invasive prenatal diagnoses |
| CA2893817C (en) * | 2012-12-13 | 2021-03-09 | Synthetic Genomics, Inc. | Peg-mediated assembly of nucleic acid molecules |
| EP3597774A1 (en) | 2013-03-13 | 2020-01-22 | Sequenom, Inc. | Primers for dna methylation analysis |
| WO2014145138A2 (en) * | 2013-03-15 | 2014-09-18 | Arnold Lyle J | Methods for amplification of nucleic acids utilizing clamp oligonuleotides |
| JP2016512696A (ja) * | 2013-03-15 | 2016-05-09 | アーノルド, ライル, ジェイ.ARNOLD, Lyle, J. | アセンブラ配列を利用する断片化された標的核酸の増幅方法 |
| WO2015021460A1 (en) * | 2013-08-09 | 2015-02-12 | Luminex Corporation | Probes for improved melt discrimination and multiplexing in nucleic acid assays |
| EP3736344A1 (en) | 2014-03-13 | 2020-11-11 | Sequenom, Inc. | Methods and processes for non-invasive assessment of genetic variations |
| EP4512908A3 (en) * | 2014-05-19 | 2025-03-26 | William Marsh Rice University | Allele-specific amplification using a composition of overlapping non-allele-specific primer and allele-specific blocker oligonucleotides |
| CN104131070A (zh) * | 2014-05-22 | 2014-11-05 | 苏州工业园区为真生物医药科技有限公司 | 一种对基因甲基化检测的方法 |
| US10889852B2 (en) | 2014-10-20 | 2021-01-12 | Commonwealth Scientific And Industrial Research Organisation | Genome methylation analysis |
| CN105331727B (zh) * | 2015-12-01 | 2019-09-03 | 湖南宏雅基因技术有限公司 | 一种人外周血循环肿瘤DNA中septin 9基因甲基化的检测试剂盒 |
| GB201612011D0 (en) | 2016-07-11 | 2016-08-24 | Glaxosmithkline Ip Dev Ltd | Novel processes for the production of oligonucleotides |
| JP6999645B2 (ja) * | 2016-08-02 | 2022-02-04 | エフ．ホフマン－ラ ロシュ アーゲー | 核酸の増幅及び検出／定量の効率を改良するためのヘルパーオリゴヌクレオチド |
| CN118562942A (zh) | 2016-09-06 | 2024-08-30 | 合成Dna技术公司 | Ngs文库浓度的标准化 |
| DE102016120124B8 (de) * | 2016-10-21 | 2018-08-23 | Gna Biosolutions Gmbh | Verfahren zum Durchführen einer Polymerase-Kettenreaktion und Vorrichtung zum Ausführen des Verfahrens |
| US10787699B2 (en) * | 2017-02-08 | 2020-09-29 | Microsoft Technology Licensing, Llc | Generating pluralities of primer and payload designs for retrieval of stored nucleotides |
| EP3530753A1 (de) * | 2018-02-26 | 2019-08-28 | AGCT GmbH | Verfahren zur primer-verlängerungsreaktion mit verbesserter spezifität |
| EP3530754A1 (de) * | 2018-02-26 | 2019-08-28 | AGCT GmbH | Verfahren zur amplifikation einer nukleinsäure mit verbesserter spezifität |
| CN112074612A (zh) * | 2018-02-26 | 2020-12-11 | Agct有限公司 | 一种具有更高特异性的核酸扩增方法 |
| US20210071243A1 (en) * | 2018-02-26 | 2021-03-11 | Agct Gmbh | Method for primer extension reaction with improved specificity |
| US20210071244A1 (en) * | 2018-02-28 | 2021-03-11 | Agct Gmbh | Method for the amplification of a nucleic acid with improved specificity |
| CN114075595B (zh) * | 2021-11-22 | 2023-11-17 | 上海交通大学 | 甲基化检测组合物、试剂盒及方法 |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6335184B1 (en) * | 1993-07-23 | 2002-01-01 | Bio-Rad Laboratories, Inc. | Linked linear amplification of nucleic acids |
| US6057134A (en) * | 1996-10-07 | 2000-05-02 | Ambion, Inc. | Modulating the efficiency of nucleic acid amplification reactions with 3' modified oligonucleotides |
| DE69827060T2 (de) * | 1997-03-20 | 2005-03-24 | F. Hoffmann-La Roche Ag | Modifizierte Primer |
| US7205129B1 (en) * | 2000-02-28 | 2007-04-17 | Qiagen Gmbh | Method for reducing artifacts in nucleic acid amplification |
| EP1420069A4 (en) * | 2001-08-20 | 2005-11-02 | Takara Bio Inc | nucleic acid amplification |
| DE60323611D1 (de) * | 2002-03-01 | 2008-10-30 | Integrated Dna Tech Inc | Polynomiale amplifikation von nukleinsäuren |
| US7629152B2 (en) * | 2002-03-01 | 2009-12-08 | Integrated Dna Technologies, Inc. | Methods for amplifying polymeric nucleic acids |
| US20050153333A1 (en) * | 2003-12-02 | 2005-07-14 | Sooknanan Roy R. | Selective terminal tagging of nucleic acids |
| US20060014183A1 (en) * | 2004-06-10 | 2006-01-19 | Pfundheller Henrik M | Extendable probes |
| US8501403B2 (en) | 2006-03-28 | 2013-08-06 | Commonwealth Scientific And Industrial Research Organisation | Amplification of DNA fragments |
| WO2008064687A1 (en) * | 2006-11-27 | 2008-06-05 | Fluimedix | Fidelity enhanced allele specific amplification method |
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2008
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- 2008-10-03 AU AU2008307153A patent/AU2008307153B2/en not_active Ceased
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- 2008-10-03 ES ES08800110.2T patent/ES2460896T3/es active Active
- 2008-10-03 CN CN200880119335.8A patent/CN101889096B/zh not_active Expired - Fee Related
- 2008-10-03 WO PCT/AU2008/001475 patent/WO2009043112A1/en not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| JP5535921B2 (ja) | 2014-07-02 |
| WO2009043112A1 (en) | 2009-04-09 |
| CN101889096A (zh) | 2010-11-17 |
| CN101889096B (zh) | 2015-10-21 |
| EP2207896A1 (en) | 2010-07-21 |
| US8455197B2 (en) | 2013-06-04 |
| JP2010539956A (ja) | 2010-12-24 |
| AU2008307153B2 (en) | 2015-03-12 |
| US20110033851A1 (en) | 2011-02-10 |
| EP2207896B1 (en) | 2014-01-29 |
| EP2207896A4 (en) | 2010-11-10 |
| AU2008307153A1 (en) | 2009-04-09 |
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