JP5457184B2 - Pcr阻害物質の存在下におけるdna増幅のためのtaqポリメラーゼ変異体酵素の使用 - Google Patents
Pcr阻害物質の存在下におけるdna増幅のためのtaqポリメラーゼ変異体酵素の使用 Download PDFInfo
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- JP5457184B2 JP5457184B2 JP2009528513A JP2009528513A JP5457184B2 JP 5457184 B2 JP5457184 B2 JP 5457184B2 JP 2009528513 A JP2009528513 A JP 2009528513A JP 2009528513 A JP2009528513 A JP 2009528513A JP 5457184 B2 JP5457184 B2 JP 5457184B2
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
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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
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Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US82569206P | 2006-09-14 | 2006-09-14 | |
| US60/825,692 | 2006-09-14 | ||
| PCT/US2007/078571 WO2008034110A2 (en) | 2006-09-14 | 2007-09-14 | Use of taq polymerase mutant enzymes for dna amplification in the presence of pcr inhibitors |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2013209506A Division JP2014054254A (ja) | 2006-09-14 | 2013-10-04 | Pcr阻害物質の存在下におけるdna増幅のためのtaqポリメラーゼ変異体酵素の使用 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2010503413A JP2010503413A (ja) | 2010-02-04 |
| JP2010503413A5 JP2010503413A5 (https=) | 2010-11-04 |
| JP5457184B2 true JP5457184B2 (ja) | 2014-04-02 |
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Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
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| JP2009528513A Expired - Fee Related JP5457184B2 (ja) | 2006-09-14 | 2007-09-14 | Pcr阻害物質の存在下におけるdna増幅のためのtaqポリメラーゼ変異体酵素の使用 |
| JP2013209506A Pending JP2014054254A (ja) | 2006-09-14 | 2013-10-04 | Pcr阻害物質の存在下におけるdna増幅のためのtaqポリメラーゼ変異体酵素の使用 |
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| JP (2) | JP5457184B2 (https=) |
| AU (1) | AU2007296180A1 (https=) |
| CA (1) | CA2664043A1 (https=) |
| WO (1) | WO2008034110A2 (https=) |
Families Citing this family (31)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7462475B2 (en) | 2004-05-20 | 2008-12-09 | Dna Poleymerase Technology, Inc. | Use of whole blood in PCR reactions |
| US20090305345A1 (en) | 2006-10-23 | 2009-12-10 | Medical Research Council | Polymerase |
| JP2011097828A (ja) * | 2008-07-09 | 2011-05-19 | Beckman Coulter Inc | 全血試料の前処理方法及び核酸増幅方法 |
| WO2010062777A2 (en) * | 2008-11-03 | 2010-06-03 | Kapabiosystems | Modified type a dna polymerases |
| FI20095729A0 (fi) * | 2009-06-26 | 2009-06-26 | Finnzymes Oy | Menetelmä deoksiribonukleiinihappojen kvantitatiiviseen PCR-monistukseen PCR-inhibiittoreita sisältävästä näytteestä |
| WO2011014885A1 (en) * | 2009-07-31 | 2011-02-03 | Agilent Technologies, Inc. | Thermostable type-a dna polymerase mutants with increased polymerization rate and resistance to inhibitors |
| FI20096013A7 (fi) * | 2009-10-02 | 2011-04-03 | Finnzymes Oy | Menetelmä reaktioseoksen valmistelemiseksi ja tähän liittyvät tuotteet |
| JP5926248B2 (ja) | 2010-06-18 | 2016-05-25 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | 増大した3’末端ミスマッチ識別能を有するdnaポリメラーゼ |
| WO2011157432A1 (en) | 2010-06-18 | 2011-12-22 | Roche Diagnostics Gmbh | Dna polymerases with increased 3'-mismatch discrimination |
| EP2582806B1 (en) | 2010-06-18 | 2015-12-16 | Roche Diagnostics GmbH | Dna polymerases with increased 3'-mismatch discrimination |
| WO2011157434A1 (en) | 2010-06-18 | 2011-12-22 | Roche Diagnostics Gmbh | Dna polymerases with increased 3'-mismatch discrimination |
| US8759062B2 (en) | 2010-06-18 | 2014-06-24 | Roche Molecular Systems, Inc. | DNA polymerases with increased 3′- mismatch discrimination |
| WO2011157436A1 (en) | 2010-06-18 | 2011-12-22 | Roche Diagnostics Gmbh | Dna polymerases with increased 3'-mismatch discrimination |
| ES2536252T3 (es) | 2010-06-18 | 2015-05-21 | F. Hoffmann-La Roche Ag | ADN polimerasas con diferenciación de desapareamientos 3' aumentada |
| CA2802239C (en) | 2010-06-18 | 2016-08-23 | F. Hoffmann-La Roche Ag | Dna polymerases with increased 3'-mismatch discrimination |
| WO2012097318A2 (en) | 2011-01-14 | 2012-07-19 | Kap Abiosystems | Modified dna polymerases for improved amplification |
| US8765435B2 (en) | 2011-02-15 | 2014-07-01 | Roche Molecular Systems, Inc. | DNA polymerases with increased 3′-mismatch discrimination |
| JP5847200B2 (ja) | 2011-02-15 | 2016-01-20 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | 増大した3’末端ミスマッチ識別能を有するdnaポリメラーゼ |
| ES2561885T3 (es) | 2011-04-11 | 2016-03-01 | F. Hoffmann-La Roche Ag | ADN polimerasas de actividad mejorada |
| GB201113430D0 (en) | 2011-08-03 | 2011-09-21 | Fermentas Uab | DNA polymerases |
| US11208636B2 (en) | 2011-08-10 | 2021-12-28 | Life Technologies Corporation | Polymerase compositions, methods of making and using same |
| US20130040365A1 (en) | 2011-08-10 | 2013-02-14 | Life Technologies Corporation | Polymerase compositions, methods of making and using same |
| ES2569723T3 (es) | 2011-12-08 | 2016-05-12 | F. Hoffmann-La Roche Ag | Polimerasas de DNA con actividad mejorada |
| ES2668448T3 (es) | 2011-12-08 | 2018-05-18 | F. Hoffmann-La Roche Ag | ADN polimerasas con actividad mejorada |
| CN103998603B (zh) | 2011-12-08 | 2016-05-25 | 霍夫曼-拉罗奇有限公司 | 具有改进活性的dna聚合酶 |
| EP3366769B1 (en) | 2012-10-16 | 2021-02-17 | DNA Polymerase Technology, Inc. | Inhibition-resistant polymerases |
| US9758773B2 (en) * | 2014-02-14 | 2017-09-12 | Agilent Technologies, Inc. | Thermostable type-A DNA polymerase mutant with increased resistance to inhibitors in blood |
| SG11201700456UA (en) | 2014-07-21 | 2017-02-27 | Agency Science Tech & Res | Silica coating on nanoparticles |
| US11091745B2 (en) | 2015-05-12 | 2021-08-17 | Dna Polymerase Technology, Inc. | Mutant polymerases and uses thereof |
| WO2021242041A1 (ko) * | 2020-05-29 | 2021-12-02 | (주)제노텍 | 유전자 변이에 대한 구분성이 향상된 dna 중합효소 변이체 |
| JP7393070B1 (ja) * | 2023-08-04 | 2023-12-06 | 株式会社ゴーフォトン | Pcr方法 |
Family Cites Families (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU8906091A (en) * | 1990-10-05 | 1992-04-28 | Wayne M. Barnes | Thermostable dna polymerase |
| EP0590327B1 (en) * | 1992-09-11 | 2003-04-09 | F. Hoffmann-La Roche Ag | Detection of nucleic acids in blood |
| US5436149A (en) * | 1993-02-19 | 1995-07-25 | Barnes; Wayne M. | Thermostable DNA polymerase with enhanced thermostability and enhanced length and efficiency of primer extension |
| US6818431B1 (en) * | 1995-10-18 | 2004-11-16 | Shanghai Mendel Dna Center Co. Ltd. | DNA polymerase having ability to reduce innate selective discrimination against fluorescent dye-labeled dideoxynucleotides |
| GB0022458D0 (en) | 2000-09-13 | 2000-11-01 | Medical Res Council | Directed evolution method |
| US20060234227A1 (en) | 2002-05-14 | 2006-10-19 | Alexei Slesarev | Helix-hairpin-helix motifs to manipulate properties of dna processing enzymes |
| US20040161767A1 (en) * | 2002-06-28 | 2004-08-19 | Baldwin Brett R. | Detection and quantification of aromatic oxygenase genes by real-time PCR |
| US7417133B2 (en) | 2004-02-27 | 2008-08-26 | Institut Pasteur | Methods for obtaining thermostable enzymes, DNA polymerase I variants from Thermus aquaticus having new catalytic activities, methods for obtaining the same, and applications of the same |
| US20050260606A1 (en) * | 2004-05-20 | 2005-11-24 | Kermekchiev Milko B | Use of whole blood in PCR reactions |
| US7462475B2 (en) * | 2004-05-20 | 2008-12-09 | Dna Poleymerase Technology, Inc. | Use of whole blood in PCR reactions |
| US20070048748A1 (en) | 2004-09-24 | 2007-03-01 | Li-Cor, Inc. | Mutant polymerases for sequencing and genotyping |
| US20070111234A1 (en) * | 2005-09-12 | 2007-05-17 | Christian Birkner | Detection of biological DNA |
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2007
- 2007-09-14 CA CA002664043A patent/CA2664043A1/en not_active Abandoned
- 2007-09-14 WO PCT/US2007/078571 patent/WO2008034110A2/en not_active Ceased
- 2007-09-14 AU AU2007296180A patent/AU2007296180A1/en not_active Abandoned
- 2007-09-14 JP JP2009528513A patent/JP5457184B2/ja not_active Expired - Fee Related
- 2007-09-14 EP EP07842556A patent/EP2061894A4/en not_active Withdrawn
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- 2013-10-04 JP JP2013209506A patent/JP2014054254A/ja active Pending
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|---|---|
| WO2008034110A9 (en) | 2013-09-12 |
| AU2007296180A1 (en) | 2008-03-20 |
| WO2008034110A2 (en) | 2008-03-20 |
| US20150368624A1 (en) | 2015-12-24 |
| CA2664043A1 (en) | 2008-03-20 |
| JP2010503413A (ja) | 2010-02-04 |
| WO2008034110A3 (en) | 2008-12-24 |
| US20110027832A1 (en) | 2011-02-03 |
| US9796965B2 (en) | 2017-10-24 |
| JP2014054254A (ja) | 2014-03-27 |
| EP2061894A4 (en) | 2010-05-05 |
| EP2061894A2 (en) | 2009-05-27 |
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