DK2002017T3 - Højkapacitet-detektering af molekylære markører baseret på restriktionsfragmenter - Google Patents
Højkapacitet-detektering af molekylære markører baseret på restriktionsfragmenter Download PDFInfo
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- DK2002017T3 DK2002017T3 DK07747276.9T DK07747276T DK2002017T3 DK 2002017 T3 DK2002017 T3 DK 2002017T3 DK 07747276 T DK07747276 T DK 07747276T DK 2002017 T3 DK2002017 T3 DK 2002017T3
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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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
- C12Q1/6855—Ligating adaptors
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6853—Nucleic acid amplification reactions using modified primers or templates
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- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
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Claims (18)
1. Fremgangsmåde til identifikation af nærværet eller fraværet af restriktionsfragmenter i en prøve, omfattende trinene: (a) tilvejebringelse af to eller flere nukleinsyreprøver; (b) fordøjelse af hver nukleinsyreprøve med mindst én restriktionsendonuklease til opnåelse af et sæt af restriktionsfragmenter; (c) tilvejebringelse af dobbeltstrengede, syntetiske adaptere indeholdende - en 5'-primer-komplementær sekvens, - en prøvespecifik identifikationssektion, - ved mindst én ende der kan ligeres til den stumpe eller udragende ende af et restriktionsfragment; (d) ligering af de dobbeltstrengede, syntetiske adaptere til restriktionsfragmenterne i sættet til tilvejebringelse af et sæt af adapter-ligerede restriktionsfragmenter; (e) eventuelt amplifikation af sættet af adapter-ligerede restriktionsfragmenter med én eller flere primere, der er i det mindste komplementære til: - den 5'-primer-komplementære sekvens i adapteren, - den prøvespecifikke identifikationssektion af adapteren og - eventuelt en sektion af adapteren, der er komplementær til de udragende rester af genkendelsessekvensen af den i det mindste ene restriktionsendonuklease, til tilvejebringelse af amplificerede adapter-ligerede restriktionsfragmenter (am-plikoner); (f) bestemmelse af sekvensen af i det mindste den prøvespecifikke identifikationssektion, resterne af genkendelsessekvensen af den i det mindste ene restriktionsendonuklease og en del af sekvensen af restriktionsfragmentet beliggende stødende op til den adapter-afledte sekvens eller stødende op til resterne af genkendelsessekvensen af den i det mindste ene restriktionsendonuklease, (g) sammenligning af to eller flere prøver for nærværet eller fraværet af adapter-ligerede restriktionsfragmenter; (h) identifikation af nærværet eller fraværet af adapter-ligerede restriktionsfragmenter i prøven.
2. Fremgangsmåde ifølge krav 1, ved hvilken restriktionsfragmenterne er molekylære markører.
3. Fremgangsmåde ifølge krav 2, ved hvilken de molekylære markører er AFLP- markører.
4. Fremgangsmåde ifølge krav 1, ved hvilken to eller flere prøver kombineres i en pool efter trinet til ligering af adapterne.
5. Fremgangsmåde ifølge krav 4, ved hvilken der for hver prøve i poolen anvendes en prøvespecifik identifikator, der adskiller sig fra de andre prøvespecifikke identifikatorer i poolen.
6. Fremgangsmåde ifølge krav 1, ved hvilken primerne indeholder ét eller flere selektive nukleotider ved 3’-enden.
7. Fremgangsmåde ifølge krav 1, ved hvilken restriktionsendonukleasen er en type Il-restriktionsendonuklease.
8. Fremgangsmåde ifølge krav 1, ved hvilken restriktionsendonukleasen er en type Ils-restriktionsendonuklease.
9. Fremgangsmåde ifølge krav 1, ved hvilken der anvendes to eller flere re-striktionsendonukleaser.
10. Fremgangsmåde ifølge krav 1, ved hvilken sekventeringen udføres ved hjælp af højkapacitet-sekventering.
11. Fremgangsmåde ifølge krav 10, ved hvilken højkapacitet-sekventeringen udføres på en fast bærer.
12. Fremgangsmåde ifølge krav 10, ved hvilken højkapacitet-sekventeringen er baseret på sekventering-ved-syntese.
13. Fremgangsmåde ifølge krav 10, ved hvilken højkapacitet-sekventeringen omfatter trinene: - annealing af amplikonerne eller de adapter-ligerede restriktionsfragmenter til perler, hvor hver perle anneales med et enkelt adapter-ligeret restriktionsfragment eller amplikon; - emulgering af perlerne i vand-i-olie-mikroreaktorer, idet hver vand-i-olie-mikro-reaktor indeholder en enkelt perle; - udførelse af emulsions-PCR til amplifikation af adapter-ligeret restriktionsfragment eller amplikoner; - eventuelt udvælgelse/berigelse af perler indeholdende amplificerede amplikoner; - påfyldning af perlerne i brønde, hvor hver brønd indeholder en enkelt perle; og - frembringelse af et pyrophosphat-signal.
14. Fremgangsmåde ifølge krav 10, ved hvilken højkapacitet-sekventeringen omfatter trinene: - annealing af de adapter-ligerede restriktionsfragmenter eller amplikoner til en overflade indeholdende henholdsvis en første og en anden primer eller en første og anden primerbindingssekvens, - udførelse af bro-amplifikation til tilvejebringelse af klynger af amplificerede adapter-ligerede restriktionsfragmenter eller amplificerede amplikoner, - bestemmelse af nukleotidsekvensen af de amplificerede adapter-ligerede restriktionsfragmenter eller amplificerede amplikoner under anvendelse af mærkede, reversible terminatornukleotider.
15. Fremgangsmåde ifølge krav 1, ved hvilken identifikatoren har en længde fra 4 till 6 bp, fortrinsvis fra 4 till 0 bp, mere foretrukket fra 4 til 8 bp, mest foretrukket fra 4 til 6 bp.
16. Fremgangsmåde ifølge krav 15, ved hvilken identifikatoren ikke indeholder 2 eller flere identiske, konsekutive baser.
17. Fremgangsmåde ifølge krav 15, ved hvilken de tilsvarende identifikatorer for to eller flere prøver indeholder mindst to forskellige nukleotider.
18. Anvendelse af fremgangsmåden som defineret i krav 1-17 til identifikation af molekylære markører, til genotypebestemmelse, bulksegregeringsanalyse, genetisk kortlægning, markør-assisteret tilbagekrydsning, kortlægning af locus for kvantitative egenskaber, kortlægning af bindingsuligevægt.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US78870606P | 2006-04-04 | 2006-04-04 | |
| US88005207P | 2007-01-12 | 2007-01-12 | |
| PCT/NL2007/000094 WO2007114693A2 (en) | 2006-04-04 | 2007-04-04 | High throughput detection of molecular markers based on aflp and high throughput sequencing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2002017T3 true DK2002017T3 (da) | 2015-09-07 |
Family
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Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK07747276.9T DK2002017T3 (da) | 2006-04-04 | 2007-04-04 | Højkapacitet-detektering af molekylære markører baseret på restriktionsfragmenter |
| DK17163116.1T DK3239304T3 (da) | 2006-04-04 | 2007-04-04 | Højgennemløbsdetektering af molekylære markører baseret på AFLP og sekventering med højt gennemløb |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK17163116.1T DK3239304T3 (da) | 2006-04-04 | 2007-04-04 | Højgennemløbsdetektering af molekylære markører baseret på AFLP og sekventering med højt gennemløb |
Country Status (8)
| Country | Link |
|---|---|
| US (6) | US20090253581A1 (da) |
| EP (3) | EP2963127B1 (da) |
| JP (2) | JP5389638B2 (da) |
| DK (2) | DK2002017T3 (da) |
| ES (3) | ES2545264T3 (da) |
| PL (1) | PL2963127T3 (da) |
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2007
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- 2007-04-04 ES ES07747276.9T patent/ES2545264T3/es active Active
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Also Published As
| Publication number | Publication date |
|---|---|
| JP2009536817A (ja) | 2009-10-22 |
| WO2007114693A3 (en) | 2007-12-21 |
| US20180291439A1 (en) | 2018-10-11 |
| ES2645661T3 (es) | 2017-12-07 |
| HK1244301A1 (en) | 2018-08-03 |
| US20120135871A1 (en) | 2012-05-31 |
| HK1219761A1 (en) | 2017-04-13 |
| US20200181694A1 (en) | 2020-06-11 |
| EP2002017B1 (en) | 2015-06-10 |
| PT2963127T (pt) | 2017-10-06 |
| EP3239304B1 (en) | 2020-08-19 |
| EP2963127B1 (en) | 2017-08-16 |
| EP2963127A1 (en) | 2016-01-06 |
| EP2002017A2 (en) | 2008-12-17 |
| WO2007114693A2 (en) | 2007-10-11 |
| DK3239304T3 (da) | 2020-10-26 |
| US20120202698A1 (en) | 2012-08-09 |
| US20140303007A1 (en) | 2014-10-09 |
| US20090253581A1 (en) | 2009-10-08 |
| US10023907B2 (en) | 2018-07-17 |
| ES2545264T3 (es) | 2015-09-09 |
| PL2963127T3 (pl) | 2018-01-31 |
| EP3239304A1 (en) | 2017-11-01 |
| ES2829295T3 (es) | 2021-05-31 |
| JP2013215212A (ja) | 2013-10-24 |
| JP5389638B2 (ja) | 2014-01-15 |
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