DK2857524T3 - Apparat til detektering af enkelt-nukleotider - Google Patents
Apparat til detektering af enkelt-nukleotider Download PDFInfo
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- DK2857524T3 DK2857524T3 DK14194399.3T DK14194399T DK2857524T3 DK 2857524 T3 DK2857524 T3 DK 2857524T3 DK 14194399 T DK14194399 T DK 14194399T DK 2857524 T3 DK2857524 T3 DK 2857524T3
- Authority
- DK
- Denmark
- Prior art keywords
- nucleic acid
- microfluidic
- stranded
- micro
- nucleotide
- Prior art date
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Classifications
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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
- C12Q1/6869—Methods for sequencing
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
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- Hematology (AREA)
- Clinical Laboratory Science (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Claims (7)
- PATENTKRAY1. Mikrofluidisk apparat til nukleinsyre-sekvensering, kendetegnet ved, at apparatet omfatter: En første zone, der omfatter et fastholdested, som rummes deri, og hvortil nukleinsyren, der skal sekvenseres, er fastholdt; et første mikrofluidisk rørsystem til foranledning af, at et første vandigt pyrophosphorolyse-medium strømmer hen over fastholdelsesstedet, hvorved det gøres muligt for nukleinsyren progressivt at blive pyrophosphorolyseret til dens bestanddele af nukleotid-triphosphater, og et andet mikrofluidisk rørsystem til fjernelse af et strømmende andet medium, som indeholder nukleotid-triphosphateme fra fastholdelses stedets omgivelser; et middel til skabelse af mikro-smådråber af det andet vandige medium i et bærer-opløsningsmiddel, som ikke er blandbart med vand, hvorved i det mindste noget af dette indeholder et af nukleotid-triphosphateme ned-strøms for den første zone; - kontaktmidler, som er indrettet til, successivt i det andet vandige medium, hvorfra mikro-smådråbeme tilvejebringes, at indføre et eller flere elementer blandt: en pyro-phosphatase, en ligase, en polymerase, et capture-system, etiketteret med fluorophoner i en ikke-detekterbar tilstand, selektivt for i det mindste et af nukleotid-triphosphateme, der er afledt fra nukleinsyren, samt en exonuklease eller polymerase med exonuklease-aktivitet med henblik på frigivelse af detekterbare fluorophorer fra et hvilket som helst anvendt capture-system ved exonukleolyse; et arrangement af mikro-smådråbe-fastholdesteder, på hvilke hver mikro-smådråbe inkuberes, indtil der optræder detekterbar fluorescens; tredje mikrofluidisk rørsystem til levering af hver mikro-smådråbe efter tur til et af fastholdestedeme; - et fluorescens-detekteringssystem, der omfatter en lyskilde, som er egnet til efter tur at ramme hvert fastholdested efter inkubation, og en detektor til detektering af fluorescens, som udsendes derfra.
- 2. Mikrofluidisk apparat til nukleinsyre-sekvensering ifølge krav 1, kendetegnet ved, at mikro-smådråbernes diameter er mindre end 50 μ·
- 3. Mikrofluidisk apparat til nukleinsyre-sekvensering ifølge krav 2, kendetegnet ved, at mikro-smådråbernes diameter ligger i intervallet mellem 2 og 20 μ.
- 4. Mikrofluidisk apparat til nukleinsyre-sekvensering ifølge et hvilket som helst af de foregående krav, kendetegnet ved, at kontaktmidlet omfatter en injektor eller en små-dråbe-udskiller.
- 5. Mikrofluidisk apparat til nukleinsyre-sekvensering ifølge et hvilket som helst af de foregående krav, kendetegnet ved, at lyskilden omfatter en laser.
- 6. Mikrofluidisk apparat til nukleinsyre-sekvensering ifølge et hvilket som helst af de foregående krav, ydermere omfattende en computer til analysering af outputtet fra detektoren med henblik på rekonstruktion af nukleinsyrens oprindelige basissekvens.
- 7. Anvendelse af det mikrofluidiske apparat til nukleinsyre-sekvensering ifølge et hvilket som helst af de foregående krav med henblik på sekvensering af DNA eller RNA.
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| EP14718147.3A EP2828408B1 (en) | 2013-04-09 | 2014-04-09 | Single nucleotide detection method |
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| GB201402644D0 (en) | 2014-02-14 | 2014-04-02 | Base4 Innovation Ltd | Methylation detection method |
| GB201412977D0 (en) * | 2014-07-22 | 2014-09-03 | Base4 Innovation Ltd | Single nucleotide detection method |
| GB201501012D0 (en) * | 2015-01-21 | 2015-03-04 | Base4 Innovation Ltd | Improved droplet sequencing apparatus and method |
| EP3115109A1 (en) | 2015-07-06 | 2017-01-11 | Base4 Innovation Limited | Droplet sequencing device |
| EP3207982A1 (en) | 2016-02-17 | 2017-08-23 | Base4 Innovation Limited | Improved droplet sequencing apparatus and method |
| EP3211092B1 (en) | 2016-02-24 | 2019-03-27 | Base4 Innovation Ltd | Single nucleotide detection method |
| EP3269444A1 (en) | 2016-07-14 | 2018-01-17 | Base4 Innovation Ltd | Method of identifying droplets in a stack and an associated sequencer |
| JP2019532623A (ja) | 2016-09-02 | 2019-11-14 | ベース４ イノベーション リミテッド | 単一ヌクレオチド検出方法および関連プローブ |
| US20200216883A1 (en) | 2016-09-06 | 2020-07-09 | Base4 Innovation Limited | Single nucleotide detection method and associated probes |
| CN109790566A (zh) | 2016-09-20 | 2019-05-21 | 贝斯4创新公司 | 单核苷酸检测方法及相关的探针 |
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| EP4450162A3 (en) | 2017-06-21 | 2025-01-08 | Lightcast Discovery Ltd | Microdroplet manipulation device |
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| CA2155186A1 (en) * | 1993-02-01 | 1994-08-18 | Kevin M. Ulmer | Methods and apparatus for dna sequencing |
| US6268146B1 (en) * | 1998-03-13 | 2001-07-31 | Promega Corporation | Analytical methods and materials for nucleic acid detection |
| AU5723700A (en) * | 1999-05-25 | 2000-12-12 | Praelux Incorporated | Method for sequency and characterizing polymeric biomolecules using aptamers anda method for producing aptamers |
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| US20050164181A1 (en) * | 2001-11-27 | 2005-07-28 | Thomas Bricson | Nanostructure, in particular for analysing individual molecules |
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| US7476501B2 (en) | 2002-03-26 | 2009-01-13 | Intel Corporation | Methods and device for DNA sequencing using surface enhanced raman scattering (SERS) |
| JP2006507921A (ja) | 2002-06-28 | 2006-03-09 | プレジデント・アンド・フェロウズ・オブ・ハーバード・カレッジ | 流体分散のための方法および装置 |
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| ATE542921T1 (de) * | 2006-04-18 | 2012-02-15 | Advanced Liquid Logic Inc | Pyrosequenzierung auf tröpfchenbasis |
| US8389297B2 (en) * | 2006-04-18 | 2013-03-05 | Duke University | Droplet-based affinity assay device and system |
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2013
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2014
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| CN105143468B (zh) | 2017-07-07 |
| JP2016515832A (ja) | 2016-06-02 |
| EP2857524B1 (en) | 2017-01-25 |
| ES2558610T3 (es) | 2016-02-05 |
| KR101876189B1 (ko) | 2018-07-09 |
| US20180044729A1 (en) | 2018-02-15 |
| ES2620982T3 (es) | 2017-06-30 |
| AU2016259441A1 (en) | 2016-12-08 |
| CN107090492A (zh) | 2017-08-25 |
| EP2857524A1 (en) | 2015-04-08 |
| US20160040224A1 (en) | 2016-02-11 |
| AU2014252804A1 (en) | 2015-10-15 |
| CN105143468A (zh) | 2015-12-09 |
| US9828631B2 (en) | 2017-11-28 |
| GB201306444D0 (en) | 2013-05-22 |
| KR20150139959A (ko) | 2015-12-14 |
| JP2017079781A (ja) | 2017-05-18 |
| US10690689B2 (en) | 2020-06-23 |
| JP6343659B2 (ja) | 2018-06-13 |
| AU2016259441B2 (en) | 2017-03-16 |
| WO2014167323A1 (en) | 2014-10-16 |
| JP6270986B2 (ja) | 2018-01-31 |
| AU2014252804B2 (en) | 2017-01-05 |
| CA2907865C (en) | 2018-02-27 |
| CA2907865A1 (en) | 2014-10-16 |
| EP2828408B1 (en) | 2015-10-14 |
| US20180044728A1 (en) | 2018-02-15 |
| US10551399B2 (en) | 2020-02-04 |
| BR112015025762A2 (pt) | 2017-10-10 |
| EP2828408A1 (en) | 2015-01-28 |
| DK2828408T3 (da) | 2015-12-14 |
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