ES2610461T3 - Método de detección de nucleótidos únicos - Google Patents

Método de detección de nucleótidos únicos Download PDF

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ES2610461T3
ES2610461T3 ES15744290.6T ES15744290T ES2610461T3 ES 2610461 T3 ES2610461 T3 ES 2610461T3 ES 15744290 T ES15744290 T ES 15744290T ES 2610461 T3 ES2610461 T3 ES 2610461T3
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oligonucleotide
produce
probe
detectable elements
nucleic acid
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Barnaby BALMFORTH
Cameron Alexander FRAYLING
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Base4 Innovation Ltd
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Base4 Innovation Ltd
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    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813Hybridisation assays
    • C12Q1/6816Hybridisation assays characterised by the detection means
    • C12Q1/6823Release of bound markers
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    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869Methods for sequencing
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    • C12Q2521/00Reaction characterised by the enzymatic activity
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    • C12Q2521/101DNA polymerase
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    • C12Q2521/00Reaction characterised by the enzymatic activity
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    • C12Q2521/50Other enzymatic activities
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    • C12Q2525/00Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
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    • C12Q2533/00Reactions characterised by the enzymatic reaction principle used
    • C12Q2533/10Reactions characterised by the enzymatic reaction principle used the purpose being to increase the length of an oligonucleotide strand
    • C12Q2533/107Probe or oligonucleotide ligation
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    • C12Q2563/00Nucleic acid detection characterized by the use of physical, structural and functional properties
    • C12Q2563/159Microreactors, 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
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    • C12Q2565/00Nucleic acid analysis characterised by mode or means of detection
    • C12Q2565/60Detection means characterised by use of a special device
    • C12Q2565/629Detection means characterised by use of a special device being a microfluidic device

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  • Chemical & Material Sciences (AREA)
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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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  • Dispersion Chemistry (AREA)

Abstract

Un método de secuenciación de un ácido nucleico caracterizado por que incluye las etapas de (1) generar una corriente de nucleósido trifosfatos únicos mediante pirofosforolisis progresiva del ácido nucleico; (2) producir al menos una sonda usada de oligonucleótido sustancialmente bicatenario mediante reacción en presencia de una polimerasa y una ligasa, comprendiendo al menos uno de los nucleósido trifosfatos únicos un sistema de sonda correspondiente (a) un primer oligonucleótido monocatenario marcado con elementos detectables característicos en un estado indetectable y (b) un segundo y tercer oligonucleótidos monocatenarios capaces de hibridar con regiones complementarias en el primer oligonucleótido; (3) digerir la sonda utilizada con una enzima que tiene actividad exonucleolítica de doble cadena para producir los elementos detectables en un estado detectable y un cuarto oligonucleótido monocatenario que es, al menos en parte, la secuencia complementaria del primer oligonucleótido; (4) hacer reaccionar el cuarto oligonucleótido con otro primer oligonucleótido para producir un producto oligonucleotídico sustancialmente bicatenario que corresponde a la sonda utilizada; (5) repetir las etapas (3) y (4) en un ciclo y (6) detectar los elementos detectables característicos liberados en cada repetición de la etapa (3).

Description

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Claims (1)

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ES15744290.6T 2014-07-22 2015-07-22 Método de detección de nucleótidos únicos Active ES2610461T3 (es)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GBGB1412977.9A GB201412977D0 (en) 2014-07-22 2014-07-22 Single nucleotide detection method
GB201412977 2014-07-22
PCT/GB2015/052119 WO2016012789A1 (en) 2014-07-22 2015-07-22 Single nucleotide detection method

Publications (1)

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ES2610461T3 true ES2610461T3 (es) 2017-04-27

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ES15744290.6T Active ES2610461T3 (es) 2014-07-22 2015-07-22 Método de detección de nucleótidos únicos
ES16197798.8T Active ES2655319T3 (es) 2014-07-22 2015-07-22 Método de detección de nucleótidos únicos

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US (3) US9856528B2 (es)
EP (2) EP3150725B1 (es)
JP (1) JP6433575B2 (es)
KR (1) KR101754091B1 (es)
CN (1) CN106471133B (es)
AU (1) AU2015293706B2 (es)
BR (1) BR112016029548A2 (es)
CA (1) CA2951212C (es)
DK (2) DK3013987T3 (es)
ES (2) ES2610461T3 (es)
GB (1) GB201412977D0 (es)
WO (1) WO2016012789A1 (es)

Families Citing this family (19)

* Cited by examiner, † Cited by third party
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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
CN109790573A (zh) * 2016-09-06 2019-05-21 贝斯4创新公司 单核苷酸检测方法及相关的探针
WO2018054964A1 (en) 2016-09-20 2018-03-29 Base4 Innovation Limited Single nucleotide detection method and associated probes
US11920192B2 (en) 2017-05-15 2024-03-05 Lightcast Discovery Ltd Single nucleotide detection method and associated probes
KR20200019717A (ko) * 2017-06-21 2020-02-24 베이스4 이노베이션 엘티디 핵산과 같은 분자를 분석하는 방법
IL271545B2 (en) 2017-06-21 2023-09-01 Base4 Innovation Ltd Microfluidic analytical device
WO2018234445A1 (en) 2017-06-21 2018-12-27 Base4 Innovation Limited MICROGOUTLET HANDLING DEVICE
US20200283832A1 (en) * 2017-10-23 2020-09-10 Base4 Innovation Ltd Single nucleotide analytical method and associated probes
EP3793729A1 (en) 2018-05-18 2021-03-24 Lightcast Discovery Ltd Droplet manipulation device and method
WO2019243577A1 (en) 2018-06-21 2019-12-26 Base4 Innovation Limited Sequencing method using modified nucleoside polyphosphates
AU2019307835B2 (en) * 2018-07-19 2021-08-19 Biofidelity Ltd Improved polynucleotide sequence detection method
ES2841907T3 (es) 2019-02-08 2021-07-12 Lightcast Discovery Ltd Método de manipulación de microgotitas
CN110964817B (zh) * 2019-11-20 2022-09-23 江西师范大学 基于核酸外切酶Ⅲ的功能化发夹探针、组合物及提高检测Pax-5a基因灵敏度的方法
EP4081660A1 (en) * 2019-12-23 2022-11-02 Biofidelity Ltd Kits and devices
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
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GB201217770D0 (en) * 2012-10-04 2012-11-14 Base4 Innovation Ltd Biological probes and the use thereof
GB201217772D0 (en) * 2012-10-04 2012-11-14 Base4 Innovation Ltd Sequencing method
US10000802B2 (en) * 2012-10-04 2018-06-19 Base4 Innovation Ltd Sequencing method
GB201300957D0 (en) 2013-01-18 2013-03-06 Base4 Innovation Ltd Sequencing method
GB201306444D0 (en) * 2013-04-09 2013-05-22 Base4 Innovation Ltd Single nucleotide detection method
GB201306445D0 (en) 2013-04-09 2013-05-22 Base4 Innovation Ltd Single nucleotide detection method

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Publication number Publication date
US10501795B2 (en) 2019-12-10
GB201412977D0 (en) 2014-09-03
EP3150725A1 (en) 2017-04-05
BR112016029548A2 (pt) 2017-10-24
EP3013987B1 (en) 2016-11-16
ES2655319T3 (es) 2018-02-19
CA2951212C (en) 2019-01-08
JP2017522031A (ja) 2017-08-10
CA2951212A1 (en) 2016-01-28
JP6433575B2 (ja) 2018-12-05
US9856528B2 (en) 2018-01-02
AU2015293706B2 (en) 2017-11-02
CN106471133A (zh) 2017-03-01
US20180080074A1 (en) 2018-03-22
CN106471133B (zh) 2019-07-09
US20180057872A1 (en) 2018-03-01
US20170130265A1 (en) 2017-05-11
KR101754091B1 (ko) 2017-07-05
AU2015293706A1 (en) 2017-02-09
EP3013987A1 (en) 2016-05-04
DK3013987T3 (en) 2017-02-27
US10519497B2 (en) 2019-12-31
KR20170018066A (ko) 2017-02-15
WO2016012789A1 (en) 2016-01-28
EP3150725B1 (en) 2017-12-06
DK3150725T3 (en) 2018-02-26

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