CA2218188A1 - Sequencage de biopolymeres en phase solide - Google Patents
Sequencage de biopolymeres en phase solide Download PDFInfo
- Publication number
- CA2218188A1 CA2218188A1 CA 2218188 CA2218188A CA2218188A1 CA 2218188 A1 CA2218188 A1 CA 2218188A1 CA 2218188 CA2218188 CA 2218188 CA 2218188 A CA2218188 A CA 2218188A CA 2218188 A1 CA2218188 A1 CA 2218188A1
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- Prior art keywords
- sequence
- nucleic acid
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- mass
- strand
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- 150000003512 tertiary amines Chemical class 0.000 description 1
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- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 1
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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
- C12Q1/6872—Methods for sequencing involving mass spectrometry
-
- 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
- C12Q1/6874—Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Electron Tubes For Measurement (AREA)
Abstract
L'invention porte sur des procédés de détection et de séquençage de séquences d'acides nucléiques cibles et de séquences d'acides nucléiques double brin, sur des sondes d'acide nucléique, sur des sondes d'acide nucléique modifiées en masse, sur des groupes de sondes intervenant dans des procédés, et sur des trousses et systèmes contenant lesdites sondes. Les procédés efficaces consistent à hybrider les acides nucléiques ou ceux qui représentent des séquences complémentaires ou homologues de la cible à un groupe de sondes d'acide nucléique lesquelles comportent une portion monobrin, une portion facultative double-brin et une séquence variable intérieure à la portion monobrin. Les poids moléculaires des acides nucléiques hybridés de l'ensemble peuvent être déterminés par spectrométrie de masse et la séquence de la cible, peut l'être en fonction du poids moléculaire des fragments. Les acides nucléiques dont les séquences peuvent être déterminées comportent des ADN et des ARN sous forme d'échantillons biologiques, tels que des biopsies prélevées sur des patients ou des échantillons prélevés dans l'environnement. Les sondes peuvent être fixées à un support solide du type pastille d'hybridation pour faciliter la mesure des poids moléculaires et l'identification de la séquence de la cible.
Applications Claiming Priority (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US41999495A | 1995-04-11 | 1995-04-11 | |
| US42000995A | 1995-04-11 | 1995-04-11 | |
| US08/419,994 | 1995-04-11 | ||
| US08/420,009 | 1995-04-11 | ||
| US08/614,151 US6436635B1 (en) | 1992-11-06 | 1996-03-12 | Solid phase sequencing of double-stranded nucleic acids |
| US08/614,151 | 1996-03-12 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2218188A1 true CA2218188A1 (fr) | 1996-10-17 |
Family
ID=27411260
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2218188 Abandoned CA2218188A1 (fr) | 1995-04-11 | 1996-04-10 | Sequencage de biopolymeres en phase solide |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0830460A1 (fr) |
| JP (1) | JPH11503611A (fr) |
| AU (1) | AU5544696A (fr) |
| CA (1) | CA2218188A1 (fr) |
| WO (1) | WO1996032504A2 (fr) |
Families Citing this family (100)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6436635B1 (en) | 1992-11-06 | 2002-08-20 | Boston University | Solid phase sequencing of double-stranded nucleic acids |
| US5795714A (en) | 1992-11-06 | 1998-08-18 | Trustees Of Boston University | Method for replicating an array of nucleic acid probes |
| US5605798A (en) | 1993-01-07 | 1997-02-25 | Sequenom, Inc. | DNA diagnostic based on mass spectrometry |
| US6194144B1 (en) | 1993-01-07 | 2001-02-27 | Sequenom, Inc. | DNA sequencing by mass spectrometry |
| EP0679196B1 (fr) | 1993-01-07 | 2004-05-26 | Sequenom, Inc. | Sequen age d'adn par spectrometrie de masse |
| US6428955B1 (en) * | 1995-03-17 | 2002-08-06 | Sequenom, Inc. | DNA diagnostics based on mass spectrometry |
| US5830655A (en) | 1995-05-22 | 1998-11-03 | Sri International | Oligonucleotide sizing using cleavable primers |
| US6146854A (en) * | 1995-08-31 | 2000-11-14 | Sequenom, Inc. | Filtration processes, kits and devices for isolating plasmids |
| DE19543065C2 (de) * | 1995-11-09 | 2002-10-31 | Gag Bioscience Gmbh Zentrum Fu | Verfahren zur Genomanalyse und Mittel zur Durchführung des Verfahrens |
| US6051378A (en) * | 1996-03-04 | 2000-04-18 | Genetrace Systems Inc. | Methods of screening nucleic acids using mass spectrometry |
| US5777324A (en) | 1996-09-19 | 1998-07-07 | Sequenom, Inc. | Method and apparatus for maldi analysis |
| GB9622665D0 (en) * | 1996-10-31 | 1997-01-08 | Zeneca Ltd | Methods |
| US7285422B1 (en) | 1997-01-23 | 2007-10-23 | Sequenom, Inc. | Systems and methods for preparing and analyzing low volume analyte array elements |
| US6133436A (en) | 1996-11-06 | 2000-10-17 | Sequenom, Inc. | Beads bound to a solid support and to nucleic acids |
| WO1998020166A2 (fr) * | 1996-11-06 | 1998-05-14 | Sequenom, Inc. | Diagnostics de l'adn fondes sur la spectrometrie de masse |
| AU5794498A (en) | 1996-12-10 | 1998-07-03 | Genetrace Systems, Inc. | Releasable nonvolatile mass-label molecules |
| US6699668B1 (en) | 1997-01-15 | 2004-03-02 | Xzillion Gmbh & Co. | Mass label linked hybridisation probes |
| ATE374836T1 (de) * | 1997-01-15 | 2007-10-15 | Xzillion Gmbh & Co Kg | Massenmarkierte hybridisierungssonden |
| US6117634A (en) | 1997-03-05 | 2000-09-12 | The Reagents Of The University Of Michigan | Nucleic acid sequencing and mapping |
| US6197557B1 (en) | 1997-03-05 | 2001-03-06 | The Regents Of The University Of Michigan | Compositions and methods for analysis of nucleic acids |
| WO1998040520A1 (fr) * | 1997-03-14 | 1998-09-17 | Hybridon, Inc. | Methode de sequençage d'acides nucleiques modifies au moyen d'une spectrometrie de masse a transformee de fourier a ionisation par electropulverisation |
| JP2001517948A (ja) | 1997-04-01 | 2001-10-09 | グラクソ、グループ、リミテッド | 核酸配列決定法 |
| AU6846698A (en) | 1997-04-01 | 1998-10-22 | Glaxo Group Limited | Method of nucleic acid amplification |
| US7056659B1 (en) | 1997-07-11 | 2006-06-06 | Xzillion Gmbh & Co. | Characterizing nucleic acids |
| US6207370B1 (en) | 1997-09-02 | 2001-03-27 | Sequenom, Inc. | Diagnostics based on mass spectrometric detection of translated target polypeptides |
| US6268131B1 (en) * | 1997-12-15 | 2001-07-31 | Sequenom, Inc. | Mass spectrometric methods for sequencing nucleic acids |
| FR2782325B1 (fr) * | 1998-08-12 | 2002-05-24 | Proteus | Procede d'identification de sequences polynucleotidiques et/ou des proteines correspondantes a partir d'un echantillon d'acides nucleiques |
| US6723564B2 (en) * | 1998-05-07 | 2004-04-20 | Sequenom, Inc. | IR MALDI mass spectrometry of nucleic acids using liquid matrices |
| US6104028A (en) * | 1998-05-29 | 2000-08-15 | Genetrace Systems Inc. | Volatile matrices for matrix-assisted laser desorption/ionization mass spectrometry |
| US7270958B2 (en) | 1998-09-10 | 2007-09-18 | The Regents Of The University Of Michigan | Compositions and methods for analysis of nucleic acids |
| AR021833A1 (es) | 1998-09-30 | 2002-08-07 | Applied Research Systems | Metodos de amplificacion y secuenciacion de acido nucleico |
| US7332275B2 (en) | 1999-10-13 | 2008-02-19 | Sequenom, Inc. | Methods for detecting methylated nucleotides |
| DE19963536C2 (de) * | 1999-12-20 | 2003-04-10 | Epigenomics Ag | Verfahren zur Analyse von Nukleinsäuresequenzen |
| JP2001204463A (ja) * | 2000-01-27 | 2001-07-31 | Toyo Kohan Co Ltd | ヌクレオチド固定用担体 |
| WO2002055199A2 (fr) | 2000-10-30 | 2002-07-18 | Sequenom Inc | Procede et dispositif d'apport de volumes inferieurs au microlitre sur un substrat |
| AR031640A1 (es) | 2000-12-08 | 2003-09-24 | Applied Research Systems | Amplificacion isotermica de acidos nucleicos en un soporte solido |
| US20040121313A1 (en) | 2002-12-06 | 2004-06-24 | Ecker David J. | Methods for rapid detection and identification of bioagents in organs for transplantation |
| US7666588B2 (en) | 2001-03-02 | 2010-02-23 | Ibis Biosciences, Inc. | Methods for rapid forensic analysis of mitochondrial DNA and characterization of mitochondrial DNA heteroplasmy |
| US7226739B2 (en) | 2001-03-02 | 2007-06-05 | Isis Pharmaceuticals, Inc | Methods for rapid detection and identification of bioagents in epidemiological and forensic investigations |
| US20030027135A1 (en) | 2001-03-02 | 2003-02-06 | Ecker David J. | Method for rapid detection and identification of bioagents |
| US7217510B2 (en) | 2001-06-26 | 2007-05-15 | Isis Pharmaceuticals, Inc. | Methods for providing bacterial bioagent characterizing information |
| JP2003043037A (ja) * | 2001-07-27 | 2003-02-13 | Inst Of Physical & Chemical Res | ハイブリダイゼーション用基板、この基板の製造方法及び使用方法 |
| GB0202462D0 (en) * | 2002-02-04 | 2002-03-20 | Tepnel Medical Ltd | Nucleic acid analysis |
| WO2003093296A2 (fr) | 2002-05-03 | 2003-11-13 | Sequenom, Inc. | Muteines de proteines d'ancrage de kinase, leurs peptides, et procedes associes |
| CA2508726A1 (fr) | 2002-12-06 | 2004-07-22 | Isis Pharmaceuticals, Inc. | Procedes d'identification rapide de pathogenes chez l'homme et les betes |
| WO2004064972A2 (fr) | 2003-01-16 | 2004-08-05 | Hk Pharmaceuticals, Inc. | Composes de capture, collections associees et methodes d'analyse du proteome et de compositions de complexes |
| US8158354B2 (en) | 2003-05-13 | 2012-04-17 | Ibis Biosciences, Inc. | Methods for rapid purification of nucleic acids for subsequent analysis by mass spectrometry by solution capture |
| US7964343B2 (en) | 2003-05-13 | 2011-06-21 | Ibis Biosciences, Inc. | Method for rapid purification of nucleic acids for subsequent analysis by mass spectrometry by solution capture |
| US9394565B2 (en) | 2003-09-05 | 2016-07-19 | Agena Bioscience, Inc. | Allele-specific sequence variation analysis |
| US20120122103A1 (en) | 2003-09-11 | 2012-05-17 | Rangarajan Sampath | Compositions for use in identification of bacteria |
| US8546082B2 (en) | 2003-09-11 | 2013-10-01 | Ibis Biosciences, Inc. | Methods for identification of sepsis-causing bacteria |
| US8097416B2 (en) | 2003-09-11 | 2012-01-17 | Ibis Biosciences, Inc. | Methods for identification of sepsis-causing bacteria |
| US8163895B2 (en) | 2003-12-05 | 2012-04-24 | Ibis Biosciences, Inc. | Compositions for use in identification of orthopoxviruses |
| US7666592B2 (en) | 2004-02-18 | 2010-02-23 | Ibis Biosciences, Inc. | Methods for concurrent identification and quantification of an unknown bioagent |
| AU2005230936B2 (en) | 2004-03-26 | 2010-08-05 | Agena Bioscience, Inc. | Base specific cleavage of methylation-specific amplification products in combination with mass analysis |
| US7608394B2 (en) | 2004-03-26 | 2009-10-27 | Sequenom, Inc. | Methods and compositions for phenotype identification based on nucleic acid methylation |
| EP1766659A4 (fr) | 2004-05-24 | 2009-09-30 | Ibis Biosciences Inc | Spectrometrie de masse a filtration ionique selective par seuillage numerique |
| US20050266411A1 (en) | 2004-05-25 | 2005-12-01 | Hofstadler Steven A | Methods for rapid forensic analysis of mitochondrial DNA |
| US7811753B2 (en) | 2004-07-14 | 2010-10-12 | Ibis Biosciences, Inc. | Methods for repairing degraded DNA |
| GB0500996D0 (en) * | 2005-01-18 | 2005-02-23 | Delfts Diagnostic Labaratory B | Detection method and materials therefor |
| US8084207B2 (en) | 2005-03-03 | 2011-12-27 | Ibis Bioscience, Inc. | Compositions for use in identification of papillomavirus |
| WO2006094238A2 (fr) | 2005-03-03 | 2006-09-08 | Isis Pharmaceuticals, Inc. | Compositions utilisees pour identifier des virus secondaires |
| GB0514910D0 (en) | 2005-07-20 | 2005-08-24 | Solexa Ltd | Method for sequencing a polynucleotide template |
| GB0514935D0 (en) | 2005-07-20 | 2005-08-24 | Solexa Ltd | Methods for sequencing a polynucleotide template |
| JP2009502137A (ja) | 2005-07-21 | 2009-01-29 | アイシス ファーマシューティカルズ インコーポレイティッド | 核酸変種の迅速な同定および定量のための方法 |
| US9550162B2 (en) * | 2005-09-19 | 2017-01-24 | Intematix Corporation | Printing liquid solution arrays for inorganic combinatorial libraries |
| GB0522310D0 (en) | 2005-11-01 | 2005-12-07 | Solexa Ltd | Methods of preparing libraries of template polynucleotides |
| GB0524069D0 (en) | 2005-11-25 | 2006-01-04 | Solexa Ltd | Preparation of templates for solid phase amplification |
| WO2007091077A1 (fr) | 2006-02-08 | 2007-08-16 | Solexa Limited | Procédé de séquençage d'une matrice polynucléotidique |
| US8088582B2 (en) | 2006-04-06 | 2012-01-03 | Ibis Biosciences, Inc. | Compositions for the use in identification of fungi |
| EP2064332B1 (fr) | 2006-09-14 | 2012-07-18 | Ibis Biosciences, Inc. | Procédé d'amplification ciblée de génome entier pour l'identification d'agents pathogènes |
| US7754429B2 (en) | 2006-10-06 | 2010-07-13 | Illumina Cambridge Limited | Method for pair-wise sequencing a plurity of target polynucleotides |
| WO2008093098A2 (fr) | 2007-02-02 | 2008-08-07 | Illumina Cambridge Limited | Procedes pour indexer des echantillons et sequencer de multiples matrices nucleotidiques |
| EP2126132B1 (fr) | 2007-02-23 | 2013-03-20 | Ibis Biosciences, Inc. | Procédé d'analyse d'adn médico-légale rapide |
| WO2008151023A2 (fr) | 2007-06-01 | 2008-12-11 | Ibis Biosciences, Inc. | Procédés et compositions pour l'amplification par déplacement multiple d'acides nucléiques |
| US8067730B2 (en) | 2007-07-20 | 2011-11-29 | The George Washington University | Laser ablation electrospray ionization (LAESI) for atmospheric pressure, In vivo, and imaging mass spectrometry |
| US7964843B2 (en) | 2008-07-18 | 2011-06-21 | The George Washington University | Three-dimensional molecular imaging by infrared laser ablation electrospray ionization mass spectrometry |
| US8901487B2 (en) | 2007-07-20 | 2014-12-02 | George Washington University | Subcellular analysis by laser ablation electrospray ionization mass spectrometry |
| US20090180931A1 (en) | 2007-09-17 | 2009-07-16 | Sequenom, Inc. | Integrated robotic sample transfer device |
| US12060554B2 (en) | 2008-03-10 | 2024-08-13 | Illumina, Inc. | Method for selecting and amplifying polynucleotides |
| US8550694B2 (en) | 2008-09-16 | 2013-10-08 | Ibis Biosciences, Inc. | Mixing cartridges, mixing stations, and related kits, systems, and methods |
| EP2347254A2 (fr) | 2008-09-16 | 2011-07-27 | Ibis Biosciences, Inc. | Unités de traitement d'échantillons, systèmes et procédés associés |
| EP2344893B1 (fr) | 2008-09-16 | 2014-10-15 | Ibis Biosciences, Inc. | Systèmes et procédés de manipulation de microplaques |
| US8728764B2 (en) | 2008-10-02 | 2014-05-20 | Illumina Cambridge Limited | Nucleic acid sample enrichment for sequencing applications |
| US8158936B2 (en) | 2009-02-12 | 2012-04-17 | Ibis Biosciences, Inc. | Ionization probe assemblies |
| US9719083B2 (en) | 2009-03-08 | 2017-08-01 | Ibis Biosciences, Inc. | Bioagent detection methods |
| US9393564B2 (en) | 2009-03-30 | 2016-07-19 | Ibis Biosciences, Inc. | Bioagent detection systems, devices, and methods |
| EP2454000A4 (fr) | 2009-07-17 | 2016-08-10 | Ibis Biosciences Inc | Systèmes pour l'identification d'un bioagent |
| WO2011008971A1 (fr) | 2009-07-17 | 2011-01-20 | Ibis Biosciences, Inc. | Appareil de levage et de montage |
| US9416409B2 (en) | 2009-07-31 | 2016-08-16 | Ibis Biosciences, Inc. | Capture primers and capture sequence linked solid supports for molecular diagnostic tests |
| EP2462244B1 (fr) | 2009-08-06 | 2016-07-20 | Ibis Biosciences, Inc. | Compositions de base déterminées non massiques pour détection d'acide nucléique |
| CA2770389C (fr) | 2009-08-25 | 2015-12-29 | Illumina, Inc. | Procedes de selection et d?amplification de polynucleotides |
| US8182994B2 (en) | 2009-09-15 | 2012-05-22 | Illumina Cambridge Limited | Centroid markers for image analysis of high denisty clusters in complex polynucleotide sequencing |
| ES2628739T3 (es) | 2009-10-15 | 2017-08-03 | Ibis Biosciences, Inc. | Amplificación por desplazamiento múltiple |
| WO2011115840A2 (fr) | 2010-03-14 | 2011-09-22 | Ibis Biosciences, Inc. | Recherche de parasites par le biais de la recherche d'endosymbiotes |
| AU2012254985C1 (en) | 2011-05-19 | 2017-02-23 | Agena Bioscience, Inc. | Products and processes for multiplex nucleic acid identification |
| JP2014524121A (ja) | 2011-07-14 | 2014-09-18 | ザ・ジョージ・ワシントン・ユニバーシティ | レーザアブレーション・エレクトロスプレイイオン化質量分析用のプルームコリメーション |
| JP6295322B2 (ja) * | 2013-10-18 | 2018-03-14 | シージーン アイエヌシー | ｈＣＴＯを利用するＰＴＯ切断及び延長分析による固相におけるターゲット核酸配列の検出 |
| WO2016172579A1 (fr) | 2015-04-24 | 2016-10-27 | Agena Bioscience, Inc, | Procédés multiplexes permettant la détection et la quantification de variants mineurs |
| JP2018512880A (ja) | 2015-04-24 | 2018-05-24 | アジーナ バイオサイエンス， インコーポレイテッド | 少量の対立遺伝子および多型の同定ならびに定量のためのマルチプレックス法 |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5795714A (en) * | 1992-11-06 | 1998-08-18 | Trustees Of Boston University | Method for replicating an array of nucleic acid probes |
| EP0679196B1 (fr) * | 1993-01-07 | 2004-05-26 | Sequenom, Inc. | Sequen age d'adn par spectrometrie de masse |
| US5650274A (en) * | 1993-06-25 | 1997-07-22 | Hitachi, Ltd. | DNA analyzing method |
-
1996
- 1996-04-10 CA CA 2218188 patent/CA2218188A1/fr not_active Abandoned
- 1996-04-10 AU AU55446/96A patent/AU5544696A/en not_active Abandoned
- 1996-04-10 WO PCT/US1996/005136 patent/WO1996032504A2/fr not_active Ceased
- 1996-04-10 EP EP96912743A patent/EP0830460A1/fr not_active Withdrawn
- 1996-04-10 JP JP8531243A patent/JPH11503611A/ja not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| EP0830460A1 (fr) | 1998-03-25 |
| AU5544696A (en) | 1996-10-30 |
| WO1996032504A3 (fr) | 1996-11-14 |
| JPH11503611A (ja) | 1999-03-30 |
| WO1996032504A2 (fr) | 1996-10-17 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |