BR112016029548A2 - método de sequenciamento de um ácido nucleico e sistema de sonda biológico multicomponente - Google Patents

método de sequenciamento de um ácido nucleico e sistema de sonda biológico multicomponente

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Publication number
BR112016029548A2
BR112016029548A2 BR112016029548A BR112016029548A BR112016029548A2 BR 112016029548 A2 BR112016029548 A2 BR 112016029548A2 BR 112016029548 A BR112016029548 A BR 112016029548A BR 112016029548 A BR112016029548 A BR 112016029548A BR 112016029548 A2 BR112016029548 A2 BR 112016029548A2
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BR
Brazil
Prior art keywords
stranded
oligonucleotide
nucleic acid
probe system
primary
Prior art date
Application number
BR112016029548A
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English (en)
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Balmforth Barnaby
Alexander Frayling Cameron
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Base4 Innovation Ltd
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Publication date
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Publication of BR112016029548A2 publication Critical patent/BR112016029548A2/pt

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    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/10—Nucleotidyl transfering
    • C12Q2521/101—DNA polymerase
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/30—Phosphoric diester hydrolysing, i.e. nuclease
    • C12Q2521/319—Exonuclease
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2521/00—Reaction characterised by the enzymatic activity
    • C12Q2521/50—Other enzymatic activities
    • C12Q2521/501—Ligase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/125—Modifications characterised by incorporating agents resulting in resistance to degradation
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/30—Oligonucleotides characterised by their secondary structure
    • C12Q2525/307—Circular oligonucleotides
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    • C12Q2533/00—Reactions characterised by the enzymatic reaction principle used
    • C12Q2533/10—Reactions characterised by the enzymatic reaction principle used the purpose being to increase the length of an oligonucleotide strand
    • C12Q2533/107—Probe or oligonucleotide ligation
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    • C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
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    • C12Q2537/149—Sequential reactions
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    • C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
    • C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
    • C12Q2537/162—Helper probe
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    • C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
    • C12Q2563/107—Nucleic acid detection characterized by the use of physical, structural and functional properties fluorescence
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    • C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
    • C12Q2563/159—Microreactors, e.g. emulsion PCR or sequencing, droplet PCR, microcapsules, i.e. non-liquid containers with a range of different permeability's for different reaction components
    • 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
    • C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
    • C12Q2565/60—Detection means characterised by use of a special device
    • C12Q2565/629—Detection means characterised by use of a special device being a microfluidic device

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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)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Dispersion Chemistry (AREA)

Abstract

a presente invenção refere-se a um método de sequenciamento de um ácido nucleico, tal como dna ou rna. dito método compreende: (1) gerar uma corrente de nucleosídeos trifosfato únicos; (2) produzir ao menos uma sonda utilizada substancialmente de fita dupla pela reação na presença de uma polimerase e uma ligase, ao menos um dos nucleosídeos trifosfato únicos com um sistema de sonda correspondente que compreende (a) um oligonucleotídeo primário de fita simples marcado com elementos característicos detectáveis em um estado indetectável; (b) oligonucleotídeos secundário e terciário de fita simples capaz de hibridizar as regiões complementares no oligonucleotídeo primário; (3) digerir a sonda utilizada com uma enzima com atividade exonucleolítica de fita dupla; (4) reagir o oligonucleotídeo quaternário com outro oligonucleotídeo primário; (5) repetir as etapas (3) e (4) em um ciclo e (6) detectar os elementos detectáveis característicos liberado em cada repetição da etapa (3). sistemas de sonda adequados são também relatados.
BR112016029548A 2014-07-22 2015-07-22 método de sequenciamento de um ácido nucleico e sistema de sonda biológico multicomponente BR112016029548A2 (pt)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1412977.9A GB201412977D0 (en) 2014-07-22 2014-07-22 Single nucleotide detection method
PCT/GB2015/052119 WO2016012789A1 (en) 2014-07-22 2015-07-22 Single nucleotide detection method

Publications (1)

Publication Number Publication Date
BR112016029548A2 true BR112016029548A2 (pt) 2017-10-24

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Country Status (12)

Country Link
US (3) US9856528B2 (pt)
EP (2) EP3013987B1 (pt)
JP (1) JP6433575B2 (pt)
KR (1) KR101754091B1 (pt)
CN (1) CN106471133B (pt)
AU (1) AU2015293706B2 (pt)
BR (1) BR112016029548A2 (pt)
CA (1) CA2951212C (pt)
DK (2) DK3150725T3 (pt)
ES (2) ES2610461T3 (pt)
GB (1) GB201412977D0 (pt)
WO (1) WO2016012789A1 (pt)

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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
US20190203273A1 (en) 2016-09-20 2019-07-04 Base4 Innovation Limited Single nucleotide detection method and associated probes
WO2018210823A1 (en) 2017-05-15 2018-11-22 Base4 Innovation Limited Single nucleotide detection method and associated probes
KR20240017985A (ko) 2017-06-21 2024-02-08 베이스4 이노베이션 엘티디 마이크로 액적 조작 장치
AU2018288533B2 (en) 2017-06-21 2023-07-13 Lightcast Discovery Ltd Microfluidic analytical device
BR112019027764A2 (pt) * 2017-06-21 2020-07-07 Base4 Innovation Limited método para manipular uma microgotícula e método para pirofosforizar um analito de ácido nucleico
CN111263818A (zh) * 2017-10-23 2020-06-09 贝斯4创新公司 单核苷酸分析方法及相关探针
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
SG11202100163TA (en) 2018-07-19 2021-02-25 Biofidelity Ltd Improved polynucleotide sequence detection method
ES2841907T3 (es) 2019-02-08 2021-07-12 Lightcast Discovery Ltd Método de manipulación de microgotitas
CN110938690B (zh) * 2019-11-20 2022-11-01 江西师范大学 基于目标触发的酶辅助发夹探针重塑的无标记比色传感器及其应用
GB201919186D0 (en) 2019-12-23 2020-02-05 Biofidelity Ltd Simplified polynucleotide sequence detection method

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JP2575270B2 (ja) 1992-11-10 1997-01-22 浜松ホトニクス株式会社 核酸の塩基配列決定方法、単一分子検出方法、その装置及び試料の作成方法
CA2155186A1 (en) 1993-02-01 1994-08-18 Kevin M. Ulmer Methods and apparatus for dna sequencing
US6361942B1 (en) * 1998-03-24 2002-03-26 Boston Probes, Inc. Method, kits and compositions pertaining to detection complexes
WO2000071755A2 (en) * 1999-05-25 2000-11-30 Praelux Incorporated Method for sequency and characterizing polymeric biomolecules using aptamers and a method for producing aptamers
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GB201217772D0 (en) * 2012-10-04 2012-11-14 Base4 Innovation Ltd Sequencing method
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GB201217770D0 (en) * 2012-10-04 2012-11-14 Base4 Innovation Ltd Biological probes and the use thereof
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GB201306445D0 (en) * 2013-04-09 2013-05-22 Base4 Innovation Ltd Single nucleotide detection method
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Also Published As

Publication number Publication date
EP3013987A1 (en) 2016-05-04
US10501795B2 (en) 2019-12-10
KR101754091B1 (ko) 2017-07-05
AU2015293706A1 (en) 2017-02-09
EP3150725B1 (en) 2017-12-06
DK3013987T3 (en) 2017-02-27
JP2017522031A (ja) 2017-08-10
JP6433575B2 (ja) 2018-12-05
EP3150725A1 (en) 2017-04-05
AU2015293706B2 (en) 2017-11-02
ES2610461T3 (es) 2017-04-27
CN106471133A (zh) 2017-03-01
US20170130265A1 (en) 2017-05-11
CA2951212C (en) 2019-01-08
GB201412977D0 (en) 2014-09-03
US20180057872A1 (en) 2018-03-01
EP3013987B1 (en) 2016-11-16
WO2016012789A1 (en) 2016-01-28
ES2655319T3 (es) 2018-02-19
CN106471133B (zh) 2019-07-09
KR20170018066A (ko) 2017-02-15
US10519497B2 (en) 2019-12-31
CA2951212A1 (en) 2016-01-28
US9856528B2 (en) 2018-01-02
DK3150725T3 (en) 2018-02-26
US20180080074A1 (en) 2018-03-22

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