WO2003080861A1 - Sequençage de molecule unique au moyen de nucleotides marques par des phosphates - Google Patents
Sequençage de molecule unique au moyen de nucleotides marques par des phosphates Download PDFInfo
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- WO2003080861A1 WO2003080861A1 PCT/EP2003/002982 EP0302982W WO03080861A1 WO 2003080861 A1 WO2003080861 A1 WO 2003080861A1 EP 0302982 W EP0302982 W EP 0302982W WO 03080861 A1 WO03080861 A1 WO 03080861A1
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- WO
- WIPO (PCT)
- Prior art keywords
- fluorescence
- labeled
- process according
- nucleic acid
- fluorescence dye
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- 229910019142 PO4 Inorganic materials 0.000 title claims description 30
- 239000010452 phosphate Substances 0.000 title claims description 23
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 title claims description 19
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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
- C12Q1/6869—Methods for sequencing
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
L'invention concerne un procédé de séquençage d'acides nucléiques selon lequel la molécule d'acide nucléique à séquencer est dégradée de manière séquentielle en présence d'un réactif marqué par fluorescence, un nucléoside marqué par fluorescence étant formé et présentant des caractéristiques de fluorescence spécifiques d'une nucléobase. De préférence, ce procédé est mis en oeuvre par une procédure de séquençage de molécule unique qui comprend une étape de détection résolue de manière spatiale, par exemple de détection confocale. De préférence, ce procédé comprend une étape de détection confocale. L'invention concerne en outre des nouveaux colorants fluorescents spécifiques d'une nucléobase et des réactifs contenant ces colorants.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP02006713 | 2002-03-22 | ||
| EP02006713.8 | 2002-03-22 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2003080861A1 true WO2003080861A1 (fr) | 2003-10-02 |
Family
ID=28051735
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/EP2003/002982 WO2003080861A1 (fr) | 2002-03-22 | 2003-03-21 | Sequençage de molecule unique au moyen de nucleotides marques par des phosphates |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2003080861A1 (fr) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003189852A (ja) * | 2001-12-26 | 2003-07-08 | Olympus Optical Co Ltd | 塩基配列決定装置および塩基配列決定方法 |
| WO2014167324A1 (fr) * | 2013-04-09 | 2014-10-16 | Base4 Innovation Ltd | Méthode de détection de mononucléotide |
| WO2014167323A1 (fr) * | 2013-04-09 | 2014-10-16 | Base4 Innovation Ltd | Procédé de détection de nucléotides simples |
| WO2014187924A1 (fr) * | 2013-05-24 | 2014-11-27 | Illumina Cambridge Limited | Sequençage pyrophosphorolytique |
| JP2015521030A (ja) * | 2012-04-09 | 2015-07-27 | ザ・トラスティーズ・オブ・コランビア・ユニバーシティー・イン・ザ・シティー・オブ・ニューヨークＴｈｅ Ｔｒｕｓｔｅｅｓ Ｏｆ Ｃｏｌｕｍｂｉａ Ｕｎｉｖｅｒｓｉｔｙ Ｉｎ Ｔｈｅ Ｃｉｔｙ Ｏｆ Ｎｅｗ Ｙｏｒｋ | ナノ細孔の調製方法およびその使用 |
| JP2015530119A (ja) * | 2012-10-04 | 2015-10-15 | ベース４ イノベーション リミテッド | 配列決定方法 |
| CN105518152A (zh) * | 2013-06-13 | 2016-04-20 | 贝斯4创新公司 | 液滴储存方法 |
| CN106471133A (zh) * | 2014-07-22 | 2017-03-01 | 贝斯4创新公司 | 单核苷酸检测方法 |
| EP3211092A1 (fr) | 2016-02-24 | 2017-08-30 | Base4 Innovation Ltd | Procédé de détection d'un nucléotide simple |
| WO2018042028A1 (fr) | 2016-09-02 | 2018-03-08 | Base4 Innovation Limited | Procédé de détection de nucléotide unique et sondes associées |
| WO2018046521A1 (fr) | 2016-09-06 | 2018-03-15 | Base4 Innovation Limited | Procédé de détection de nucléotides simples et sondes associées |
| WO2018054964A1 (fr) | 2016-09-20 | 2018-03-29 | Base4 Innovation Limited | Procédé de détection de mononucléotides et sondes associées |
| US10000802B2 (en) | 2012-10-04 | 2018-06-19 | Base4 Innovation Ltd | Sequencing method |
| WO2018210823A1 (fr) | 2017-05-15 | 2018-11-22 | Base4 Innovation Limited | Procédé de détection de nucléotides uniques et sondes associées |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4962037A (en) * | 1987-10-07 | 1990-10-09 | United States Of America | Method for rapid base sequencing in DNA and RNA |
| WO1994018218A1 (fr) * | 1993-02-01 | 1994-08-18 | Seq, Ltd. | Procedes et appareil de sequençage de l'adn |
| WO1999013109A1 (fr) * | 1997-09-11 | 1999-03-18 | Seq, Ltd. | Procede d'analyse de sequençage d'adn |
| US6255083B1 (en) * | 1998-12-14 | 2001-07-03 | Li Cor Inc | System and methods for nucleic acid sequencing of single molecules by polymerase synthesis |
| WO2001094609A1 (fr) * | 2000-06-07 | 2001-12-13 | Li-Cor, Inc. | Nucleotides a commutation de charges |
| WO2002002795A2 (fr) * | 2000-06-30 | 2002-01-10 | Gnothis Holding Sa | Procede de sequençage multiplex |
-
2003
- 2003-03-21 WO PCT/EP2003/002982 patent/WO2003080861A1/fr not_active Application Discontinuation
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4962037A (en) * | 1987-10-07 | 1990-10-09 | United States Of America | Method for rapid base sequencing in DNA and RNA |
| WO1994018218A1 (fr) * | 1993-02-01 | 1994-08-18 | Seq, Ltd. | Procedes et appareil de sequençage de l'adn |
| WO1999013109A1 (fr) * | 1997-09-11 | 1999-03-18 | Seq, Ltd. | Procede d'analyse de sequençage d'adn |
| US6255083B1 (en) * | 1998-12-14 | 2001-07-03 | Li Cor Inc | System and methods for nucleic acid sequencing of single molecules by polymerase synthesis |
| WO2001094609A1 (fr) * | 2000-06-07 | 2001-12-13 | Li-Cor, Inc. | Nucleotides a commutation de charges |
| WO2002002795A2 (fr) * | 2000-06-30 | 2002-01-10 | Gnothis Holding Sa | Procede de sequençage multiplex |
Non-Patent Citations (3)
| Title |
|---|
| ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, vol. 175, no. 1, 1976, pages 185 - 189, ISSN: 0003-9861 * |
| DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1976, HUANG W ET AL: "TRANSPORT ATPASE OF ERYTHROCYTE MEMBRANE SENSITIVITIES OF SODIUM POTASSIUM ACTIVATED ATPASE AND POTASSIUM PHOSPHATASE ACTIVITIES TO OUABAIN", XP002227915, Database accession no. PREV197662050423 * |
| STEPHAN J ET AL: "Towards a general procedure for sequencing single DNA molecules", JOURNAL OF BIOTECHNOLOGY, ELSEVIER SCIENCE PUBLISHERS, AMSTERDAM, NL, vol. 86, no. 3, 13 April 2001 (2001-04-13), pages 255 - 267, XP004230519, ISSN: 0168-1656 * |
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|---|---|---|---|---|
| JP2003189852A (ja) * | 2001-12-26 | 2003-07-08 | Olympus Optical Co Ltd | 塩基配列決定装置および塩基配列決定方法 |
| JP2015521030A (ja) * | 2012-04-09 | 2015-07-27 | ザ・トラスティーズ・オブ・コランビア・ユニバーシティー・イン・ザ・シティー・オブ・ニューヨークＴｈｅ Ｔｒｕｓｔｅｅｓ Ｏｆ Ｃｏｌｕｍｂｉａ Ｕｎｉｖｅｒｓｉｔｙ Ｉｎ Ｔｈｅ Ｃｉｔｙ Ｏｆ Ｎｅｗ Ｙｏｒｋ | ナノ細孔の調製方法およびその使用 |
| JP2015530119A (ja) * | 2012-10-04 | 2015-10-15 | ベース４ イノベーション リミテッド | 配列決定方法 |
| CN108220146A (zh) * | 2012-10-04 | 2018-06-29 | 贝斯4创新公司 | 测序方法 |
| US10000802B2 (en) | 2012-10-04 | 2018-06-19 | Base4 Innovation Ltd | Sequencing method |
| EP3327143A1 (fr) * | 2012-10-04 | 2018-05-30 | Base4 Innovation Ltd | Procédé de séquençage |
| US9828631B2 (en) | 2013-04-09 | 2017-11-28 | Base4 Innovation Ltd | Single nucleotide detection method |
| US10480024B2 (en) | 2013-04-09 | 2019-11-19 | Base4 Innovation Ltd | Single nucleotide detection method |
| KR101876189B1 (ko) * | 2013-04-09 | 2018-07-09 | 베이스4 이노베이션 엘티디 | 단일 뉴클레오티드 검출 방법 |
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| US10551399B2 (en) | 2013-04-09 | 2020-02-04 | Base4 Innovation Ltd | Single nucleotide detection method |
| AU2014252804B2 (en) * | 2013-04-09 | 2017-01-05 | Base4 Innovation Ltd | Single nucleotide detection method |
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| JP2017079781A (ja) * | 2013-04-09 | 2017-05-18 | ベース４ イノベーション リミテッド | マイクロ流体核酸配列決定装置及びその使用 |
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| CN105143468B (zh) * | 2013-04-09 | 2017-07-07 | 贝斯4创新公司 | 单核苷酸检测方法 |
| EP2857524A1 (fr) * | 2013-04-09 | 2015-04-08 | Base4 Innovation Limited | Appareil pour la détection d'un nucléotide simple |
| AU2014270410B2 (en) * | 2013-05-24 | 2020-07-16 | Illumina Cambridge Limited | Pyrophosphorolytic sequencing |
| EP3825415A1 (fr) * | 2013-05-24 | 2021-05-26 | Illumina Cambridge Limited | Séquençage pyrophosphorolytique utilisant des nanopores |
| US11286521B2 (en) | 2013-05-24 | 2022-03-29 | Illumina Cambridge Limited | Pyrophosphorolytic sequencing |
| JP2016526876A (ja) * | 2013-05-24 | 2016-09-08 | イルミナ ケンブリッジ リミテッド | 加ピロリン酸分解式配列決定法 |
| US10450605B2 (en) | 2013-05-24 | 2019-10-22 | Illumina Cambridge Limited | Pyrophosphorolytic sequencing |
| WO2014187924A1 (fr) * | 2013-05-24 | 2014-11-27 | Illumina Cambridge Limited | Sequençage pyrophosphorolytique |
| CN105264093A (zh) * | 2013-05-24 | 2016-01-20 | 伊鲁米纳剑桥有限公司 | 焦磷酸解测序 |
| EP3351645A1 (fr) * | 2013-05-24 | 2018-07-25 | Illumina Cambridge Limited | Séquençage pyrophosphorolytique utilisant des nanopores |
| CN105518152B (zh) * | 2013-06-13 | 2018-12-18 | 贝斯4创新公司 | 液滴储存方法 |
| US10000794B2 (en) | 2013-06-13 | 2018-06-19 | Base4 Innovation Ltd | Droplet storage method |
| JP2016521558A (ja) * | 2013-06-13 | 2016-07-25 | ベース４ イノベーション リミテッド | 液滴保存方法 |
| CN105518152A (zh) * | 2013-06-13 | 2016-04-20 | 贝斯4创新公司 | 液滴储存方法 |
| CN106471133A (zh) * | 2014-07-22 | 2017-03-01 | 贝斯4创新公司 | 单核苷酸检测方法 |
| CN106471133B (zh) * | 2014-07-22 | 2019-07-09 | 贝斯4创新公司 | 单核苷酸检测方法 |
| US10501795B2 (en) | 2014-07-22 | 2019-12-10 | Base4 Innovation Ltd | Single nucleotide detection method |
| US10519497B2 (en) | 2014-07-22 | 2019-12-31 | Base4 Innovation Ltd | Single nucleotide detection method |
| US9856528B2 (en) | 2014-07-22 | 2018-01-02 | Base4 Innovation Ltd | Single nucleotide detection method |
| KR101754091B1 (ko) | 2014-07-22 | 2017-07-05 | 베이스4 이노베이션 엘티디 | 단일 뉴클레오티드 검출 방법 |
| WO2017144653A1 (fr) | 2016-02-24 | 2017-08-31 | Base4 Innovation Limited | Procédé de détection de nucléotides simples |
| EP3211092A1 (fr) | 2016-02-24 | 2017-08-30 | Base4 Innovation Ltd | Procédé de détection d'un nucléotide simple |
| WO2018042028A1 (fr) | 2016-09-02 | 2018-03-08 | Base4 Innovation Limited | Procédé de détection de nucléotide unique et sondes associées |
| WO2018046521A1 (fr) | 2016-09-06 | 2018-03-15 | Base4 Innovation Limited | Procédé de détection de nucléotides simples et sondes associées |
| WO2018054964A1 (fr) | 2016-09-20 | 2018-03-29 | Base4 Innovation Limited | Procédé de détection de mononucléotides et sondes associées |
| WO2018210823A1 (fr) | 2017-05-15 | 2018-11-22 | Base4 Innovation Limited | Procédé de détection de nucléotides uniques et sondes associées |
| US11920192B2 (en) | 2017-05-15 | 2024-03-05 | Lightcast Discovery Ltd | Single nucleotide detection method and associated probes |
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