DK3004378T3 - Pyrophosphorolytisk sekventering ved anvendelse af nanoporer - Google Patents
Pyrophosphorolytisk sekventering ved anvendelse af nanoporer Download PDFInfo
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- DK3004378T3 DK3004378T3 DK14725498.1T DK14725498T DK3004378T3 DK 3004378 T3 DK3004378 T3 DK 3004378T3 DK 14725498 T DK14725498 T DK 14725498T DK 3004378 T3 DK3004378 T3 DK 3004378T3
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- nucleic acid
- polymerase
- nanopore
- target nucleic
- nucleotide
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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
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
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- Molecular Biology (AREA)
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- Genetics & Genomics (AREA)
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- Biotechnology (AREA)
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- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Chemical Kinetics & Catalysis (AREA)
Claims (10)
1. Fremgangsmåde ti! bestemmelse af sekvensen af en målnukleinsyre, der omfatter (a) tilvejebringelse af en måinukieinsyre med to strenge: (b) etablering af kontakt mellem må!nukleinsyren og en polymerase under betingelser fil sél^entiei i ierneise af nukleotider fra den første af de to strenge ved hjaslp af pyføphøsphorolyse til derved sekventiel frembringelse af nukleotidtriphosphater med mange forskellige basedele; og (c) skelnen mellem de forskellige basedele for de sekventielt dannede nukleotidtriphosphater til derved bestemmelse af sekvensen for måinukleinsyren: hvor skelnen åf de forskellige basedele for de sekventielt dannede nukleotldiriphosphater omfatter passage af nukieotidtriphosphaterne gennem en na n opore.
2. Fremgangsmåde ifølge krav 1, hvor polymerasen er bundet til nanoporen. $; Fremgangsmåde ifølge krav 1, hvor betingelserne til sekventiel f|efnelse af nukieotider fra den ene af de to strenge ved hjælp af pyrophosphoroiyse omfatter etablering af kontakt mellem polymerasen og en pyrophosphorolytisk koncentration af pyrophosphai 4 Fremgangsmåde ifølge krav 3, hvor fremgangsmåden yderligere omfatter et trin med pause i den sekventielle frembringelse af nukieotidtriphosphaterne ved fjernelse af pyrophosphai fra kontakt med polymerasen og efterfølgende genoptagelse af den sekventielle frembringelse af nukieotidtriphosphaterne ved etablering af kontakt mellem polymerasen og pyrophosphai.
5. Fremgangsmåde Ifølge krav 1, hvor de mangeforskeiiige basedele omfatter mindst to forskellige former af basedele og højst fire forskellige former af basédéie.
6. Fremgangsmåde ifølge krav 5, hvor sekvensem der bestemmes, er længere end ire nukleotider.
7. Fremgangsmåde ifølge krav hvor basedeiene omfatter naturligt forekommende adenin. guanin, cytosin eller thymin.
3. Fremgangsmåde ifølge krav 1, hvor mindst én af basedelene omfatter en del, der er ikke-naturiigt forekommende i DMA eller RNA.
9. Apparat, der omfatter (a) en fluid-irnpermeabef barriere, der adskiller et første fluidreservoir fra et andet fiuidæservoir; (b) en nanopore, der er placeret i den fiuid-impermeabie bardere til frembringelse af en passage, hvorigennem et nukleotidtriphosphat kan passere fra det første fluidreservoir ti! det andet fiuldreservoir; og (c) en reaktionsblanding i det første fluidreservoir, hvilken reaktionsblanding omfatter en polymerase, målnukteinsyre med to strenge og en pyrophosphorofytisk koncentration af pyrpphosphat. il Q, Apparat ifølge krav 9, hvor polymerasen er bandet $1nanopørm
11. Apparat ifølge krav 9, hvor den fiuid-imperroeabSe barriere omfatter en membran, og hvor en streng af måinukieinsyren er bundet il membranen.
12. Apparat ifølge krav 11, hvor måinukieinsyren indbefatter mindst én basedej, def er ikke-naturligt forekommende i DNA eller SNA.
13. Apparat ifølge krav 9, hvor nanoporen omfatter en fastforronanopore.
14. Apparat ifølge krav 9, hvor polymerasen mangler S'-mad-S-exonukleaseaktivitet
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| US201361827175P | 2013-05-24 | 2013-05-24 | |
| PCT/EP2014/060588 WO2014187924A1 (en) | 2013-05-24 | 2014-05-22 | Pyrophosphorolytic sequencing |
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| DK3004378T3 true DK3004378T3 (da) | 2018-03-26 |
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| ES2660671T3 (es) * | 2013-05-24 | 2018-03-23 | Illumina Cambridge Limited | Secuenciado pirofosforolítico usando nanoporos |
| AU2015208919B9 (en) | 2014-01-22 | 2021-04-01 | Oxford Nanopore Technologies Limited | Method for attaching one or more polynucleotide binding proteins to a target polynucleotide |
| EP3137627B1 (en) | 2014-05-02 | 2025-07-30 | Oxford Nanopore Technologies plc | Method of improving the movement of a target polynucleotide with respect to a transmembrane pore |
| KR20250130441A (ko) | 2014-09-01 | 2025-09-01 | 브이아이비 브이지더블유 | 돌연변이체 csgg 포어 |
| US11186868B2 (en) | 2016-03-02 | 2021-11-30 | Oxford Nanopore Technologies Plc | Mutant pore |
| CN118326018B (zh) | 2016-04-06 | 2025-11-07 | 牛津纳米孔科技公开有限公司 | 突变体孔 |
| WO2018195222A1 (en) | 2017-04-19 | 2018-10-25 | Electronic Biosciences, Inc. | Nanopore/nanowell electrode enabled exonuclease sequencing |
| GB201707122D0 (en) | 2017-05-04 | 2017-06-21 | Oxford Nanopore Tech Ltd | Pore |
| CN117106037B (zh) | 2017-06-30 | 2025-07-25 | 弗拉芒区生物技术研究所 | 新颖蛋白孔 |
| GB201818216D0 (en) | 2018-11-08 | 2018-12-26 | Oxford Nanopore Tech Ltd | Pore |
| CN113195736A (zh) | 2018-11-08 | 2021-07-30 | 牛津纳米孔科技公司 | 孔 |
| JP2022529623A (ja) | 2019-04-09 | 2022-06-23 | オックスフォード ナノポール テクノロジーズ ピーエルシー | 細孔 |
| GB201907244D0 (en) | 2019-05-22 | 2019-07-03 | Oxford Nanopore Tech Ltd | Method |
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| GB202211602D0 (en) | 2022-08-09 | 2022-09-21 | Oxford Nanopore Tech Plc | Novel pore monomers and pores |
| KR20250096716A (ko) | 2022-10-28 | 2025-06-27 | 옥스포드 나노포어 테크놀로지즈 피엘씨 | 포어 단량체 및 포어 |
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| US20140349300A1 (en) | 2014-11-27 |
| CA2907423A1 (en) | 2014-11-27 |
| EP3825415B1 (en) | 2024-08-28 |
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| HK1261919A1 (en) | 2020-01-10 |
| US11286521B2 (en) | 2022-03-29 |
| US20200157619A1 (en) | 2020-05-21 |
| PT3004378T (pt) | 2018-03-06 |
| DK3351645T3 (da) | 2020-09-21 |
| EP3351645A1 (en) | 2018-07-25 |
| EP3825415A1 (en) | 2021-05-26 |
| US20220177960A1 (en) | 2022-06-09 |
| EP3351645B1 (en) | 2020-07-15 |
| JP2016526876A (ja) | 2016-09-08 |
| ES2660671T3 (es) | 2018-03-23 |
| US10450605B2 (en) | 2019-10-22 |
| EP3004378B1 (en) | 2017-12-20 |
| JP6329624B2 (ja) | 2018-05-23 |
| AU2014270410B2 (en) | 2020-07-16 |
| CN105264093B (zh) | 2018-04-10 |
| EP3004378A1 (en) | 2016-04-13 |
| CN105264093A (zh) | 2016-01-20 |
| US20180105874A1 (en) | 2018-04-19 |
| ES2994071T3 (en) | 2025-01-16 |
| ES2817425T3 (es) | 2021-04-07 |
| WO2014187924A1 (en) | 2014-11-27 |
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