DK2904114T3 - Fremgangsmåde til detektering af et enkelt-nukleotid - Google Patents

Fremgangsmåde til detektering af et enkelt-nukleotid Download PDF

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Publication number
DK2904114T3
DK2904114T3 DK13798365.6T DK13798365T DK2904114T3 DK 2904114 T3 DK2904114 T3 DK 2904114T3 DK 13798365 T DK13798365 T DK 13798365T DK 2904114 T3 DK2904114 T3 DK 2904114T3
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DK
Denmark
Prior art keywords
probe
target
detectable
nucleotide
stranded
Prior art date
Application number
DK13798365.6T
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English (en)
Inventor
Cameron Alexander Frayling
Barnaby Balmforth
Bruno Flavio Nogueira De Sousa Soares
Thomas Henry Isaac
Boris Breiner
Alessandra Natale
Michele Amasio
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Base4 Innovation Ltd
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Publication of DK2904114T3 publication Critical patent/DK2904114T3/da

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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
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • 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
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6823—Release of bound markers
    • 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)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Claims (10)

  1. PATENTKRAY
    1. Fremgangsmåde til detektering af et enkeltnukleotid-mål, kendetegnet ved, at den omfatter trinnet at fastgøre målet til en probe ved anvendelse af en polymerase og en ligase med henblik på dannelse af en anvendt probe, som helt er dobbelt-strenget, hvorved proben omfatter et ind-fangningssted, som består af et enkeltnukleotid, der er komplementært i forhold til målet, hvis ender er fastgjort til to forskellige dobbelt-strengede oligo-nukleotid-områder, hvorved i det mindste et af oligonukleotid-områderne omfatter detekterbare elementer med karakteristisk detekterings-egenskab, og hvorved de detekterbare elementer er arrangeret således på oligonukleotid-området, at detekterings-egenskaben i mindre grad er detekterbar, end når samme antal detekterbare elementer er bundet til et tilsvarende antal enkeltnu-kleotider.
  2. 2. Fremgangsmåde ifølge krav 1, kendetegnet ved, at den omfatter det ekstra trin at behandle den anvendte probe med et restriktions-enzym og en exonuklease for at frigive det/de detekterbare element/elementer i en form, i hvilken de kan detekteres.
  3. 3. Fremgangsmåde ifølge krav 2, kendetegnet ved, at den desuden omfatter trinnet at observere den detekterbare egenskab, som udvises af det/de frigivne detekterbare element(er).
  4. 4. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 3, kendetegnet ved, at det/de detekterbare element/elementer er fluoro-phorer.
  5. 5. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, kendetegnet ved, at den anvendte probe omfatter et restriktionsen-zym-genkendelsessted, der er blevet dannet ved hæftning af målet til indfang-ningsstedet.
  6. 6. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, kendetegnet ved, at målet bringes i kontakt med en blanding af fire forskellige biologiske prober, der hver omfatter et enkelt-strenget nukleotid-område, som omfatter et forskelligt nukleotid, der er udvalgt blandt (1) guanin, cytosin, adenin og thymin eller (2) guanin, cytosin, adenin og uracil.
  7. 7. Fremgangsmåde ifølge krav 6, kendetegnet ved, at hver af de fire prober omfatter et forskelligt, de-tekterbart element.
  8. 8. Fremgangsmåde ifølge krav 7, kendetegnet ved, at de forskellige, detekterbare elementer er fluoro-phorer.
  9. 9. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, kendetegnet ved, at målet omfatter eet blandt en strøm af enkelt-nukleotider, svarende til sekvensen af nukleotider i et DNA- eller RNA-prøveemne.
  10. 10. Anvendelse af en fremgangsmåde ifølge et hvilket som helst af de foregående krav med henblik på sekvensering af DNA eller RNA.
DK13798365.6T 2012-10-04 2013-10-04 Fremgangsmåde til detektering af et enkelt-nukleotid DK2904114T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1217770.5A GB201217770D0 (en) 2012-10-04 2012-10-04 Biological probes and the use thereof
PCT/GB2013/052594 WO2014053853A1 (en) 2012-10-04 2013-10-04 Biological probes and the use thereof

Publications (1)

Publication Number Publication Date
DK2904114T3 true DK2904114T3 (da) 2017-03-13

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Family Applications (1)

Application Number Title Priority Date Filing Date
DK13798365.6T DK2904114T3 (da) 2012-10-04 2013-10-04 Fremgangsmåde til detektering af et enkelt-nukleotid

Country Status (10)

Country Link
US (1) US20150275293A1 (da)
EP (1) EP2904114B1 (da)
JP (1) JP6403674B2 (da)
KR (1) KR20150060987A (da)
CN (1) CN104884635B (da)
CA (1) CA2887320A1 (da)
DK (1) DK2904114T3 (da)
ES (1) ES2613863T3 (da)
GB (1) GB201217770D0 (da)
WO (1) WO2014053853A1 (da)

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GB201217772D0 (en) * 2012-10-04 2012-11-14 Base4 Innovation Ltd Sequencing method
US10227638B2 (en) 2014-06-02 2019-03-12 Base4 Innovation Ltd. Nucleotide polymorphism 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
CN109790572A (zh) * 2016-09-02 2019-05-21 贝斯4创新公司 单核苷酸检测方法及相关探针
US20200216883A1 (en) * 2016-09-06 2020-07-09 Base4 Innovation Limited Single nucleotide detection method and associated probes
WO2018054964A1 (en) 2016-09-20 2018-03-29 Base4 Innovation Limited Single nucleotide detection method and associated probes
EP3625366A1 (en) * 2017-05-15 2020-03-25 Base4 Innovation Limited Single nucleotide detection method and associated probes
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EP3912725A1 (en) 2017-06-21 2021-11-24 Lightcast Discovery Ltd Method for investigating molecules such as nucleic acids
WO2019219905A1 (en) 2018-05-18 2019-11-21 Base4 Innovation Limited Droplet manipulation device and method
WO2019243577A1 (en) 2018-06-21 2019-12-26 Base4 Innovation Limited Sequencing method using modified nucleoside polyphosphates

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Also Published As

Publication number Publication date
KR20150060987A (ko) 2015-06-03
CN104884635A (zh) 2015-09-02
EP2904114B1 (en) 2016-11-30
JP6403674B2 (ja) 2018-10-10
EP2904114A1 (en) 2015-08-12
US20150275293A1 (en) 2015-10-01
ES2613863T3 (es) 2017-05-26
WO2014053853A1 (en) 2014-04-10
JP2015532098A (ja) 2015-11-09
GB201217770D0 (en) 2012-11-14
CN104884635B (zh) 2017-08-25
CA2887320A1 (en) 2014-04-10

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