DK2565279T3 - Effektiv basebestemmelse i sekvenseringsreaktioner - Google Patents
Effektiv basebestemmelse i sekvenseringsreaktioner Download PDFInfo
- Publication number
- DK2565279T3 DK2565279T3 DK12164915.6T DK12164915T DK2565279T3 DK 2565279 T3 DK2565279 T3 DK 2565279T3 DK 12164915 T DK12164915 T DK 12164915T DK 2565279 T3 DK2565279 T3 DK 2565279T3
- Authority
- DK
- Denmark
- Prior art keywords
- adaptor
- nucleic acid
- probe
- anchor
- sequencing
- Prior art date
Links
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
- C12Q1/6874—Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation
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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
- C12Q1/6813—Hybridisation assays
- C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
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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
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
Claims (14)
1. Fremgangsmåde til identificering af et nukleotid ved en måldetekteringsposition for en målsekvens omfattende en flerhed af detekteringspositioner, hvilken fremgangsmåde omfatter: (a) tilvejebringelse af en overflade omfattende en flerhed af concatemerer, der er immobiliseret derpå, hvor overfladen omfatter en flerhed af rumligt forskellige regioner omfattende de immobiliserede concatemerer, hvor hver concatemer omfatter en flerhed af monomerer, og hvor hver monomer omfatter: i) et måldomæne af målsekvensen omfattende et første sæt af måldetekteringspositioner; ii) en første adapter omfattende et forankringssted; b) hybridisering af en første ankersonde til forankringsstedet; c) hybridisering af en anden ankersonde til en region af målsekvensen, der er mellem det første forankringssted og måldomænet, og grænser op til begge, hvor den anden ankersonde er fuldt degenereret; d) hybridisering af en sekvenseringssonde til måldomænet, hvor sekvenseringssonden omfatter: i) en sekvens, der er komplementær til måldomænet, hvor sekvensen indbefatter et nukleotid ved en interrogationsposition, hvor nukleotidet er komplementært til et nukleotid ved en detekteringsposition; og ii) en markør, der identificerer nukleotidet ved interrogationspositionen; e) ligering af ankersondeme og sekvenseringssonden for at danne et sondeligeringsprodukt; og derefter f) detektering af sondeligeringsproduktet for derved at identificere nukleotidet.
2. Fremgangsmåde ifølge krav 1, hvor et sæt sekvenseringssonder bringes i kontakt med overfladen, hvor hver sekvenseringssonde omfatter: a) en sekvens, der er komplementær til et måldomæne af en monomer; b) et unikt nukleotid ved en første interrogationsposition; og c) en markør; hvor hver markør fra sættet svarer til det unikke nukleotid.
3. Fremgangsmåde ifølge krav 1, hvor hver monomer omfatter en flerhed af adaptere.
4. Fremgangsmåde ifølge krav 3, hvor mindst én af adapterne omfatter mindst ét Type lls-endonukleasegenkendelsessted.
5. Fremgangsmåde ifølge krav 1, hvor trin b)-e) gentages over flere cyklusser for at identificere nukleotider ved supplerende måldetekteringspositioner i det første sæt af måldetekteringspositioner, hvor der i forskellige cyklusser anvendes forskellige sekvenseringssonder, hvor de forskellige sekvenseringssonder adskiller sig ved at have et unikt nukleotid ved forskellige interrogationspositioner.
6. Fremgangsmåde ifølge krav 1, hvor den anden ankersonde omfatter mindst én terminus, der selektivt kan aktiveres for ligering.
7. Fremgangsmåde ifølge krav 1, hvor overfladen er funktionaliseret.
8. Fremgangsmåde ifølge krav 7, hvor den funktionaliserede overflade omfatter funktionelle enheder, der er udvalgt ffa gruppen bestående af aminer, silaner og hydroxylers.
9. Fremgangsmåde ifølge krav 1, hvor concatememe er immobiliseret på overfladen ved anvendelse af opfangningssonder.
10. Fremgangsmåde ifølge krav 1, hvilken fremgangsmåde endvidere omfatter fragmentering af genomisk nukleinsyre for at danne målsekvenser.
11. Fremgangsmåde ifølge krav 1, hvor målsekvensen er en genomisk nukleinsyresekvens.
12. Fremgangsmåde ifølge krav 12, hvor de genomiske nukleinsyresekvenser er humane.
13. Fremgangsmåde ifølge krav 1, hvor a) den anden ankersonde er phosphoryleret ved 5'- og 3'-termini; eller b) den første ankersonde er phosphoryleret ved dens 5'-terminus og den anden ankersonde ikke er phosphoryleret ved dens 5'- og 3'-termini.
14. Fremgangsmåde ifølge krav 1, hvor den første og anden ankersonde er ligerede i første trin for at danne et udvidet anker, og i et separat efterfølgende trin sekvenseringssonden er ligeret til det udvidede anker for at danne sondeligeringsproduktet.
Applications Claiming Priority (9)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US99248507P | 2007-12-05 | 2007-12-05 | |
US2633708P | 2008-02-05 | 2008-02-05 | |
US3591408P | 2008-03-12 | 2008-03-12 | |
US6113408P | 2008-06-13 | 2008-06-13 | |
US10258608P | 2008-10-03 | 2008-10-03 | |
US12/265,593 US7901890B2 (en) | 2007-11-05 | 2008-11-05 | Methods and oligonucleotide designs for insertion of multiple adaptors employing selective methylation |
US12/266,385 US7897344B2 (en) | 2007-11-06 | 2008-11-06 | Methods and oligonucleotide designs for insertion of multiple adaptors into library constructs |
US11619308P | 2008-11-19 | 2008-11-19 | |
EP08858697A EP2227563B1 (en) | 2007-12-05 | 2008-12-05 | Efficient base determination in sequencing reactions |
Publications (1)
Publication Number | Publication Date |
---|---|
DK2565279T3 true DK2565279T3 (da) | 2015-02-16 |
Family
ID=52465106
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DK12164915.6T DK2565279T3 (da) | 2007-12-05 | 2008-12-05 | Effektiv basebestemmelse i sekvenseringsreaktioner |
DK12191212.5T DK2610351T3 (da) | 2007-12-05 | 2008-12-05 | Effektiv basebestemmelse i sekvenseringsreaktioner |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
DK12191212.5T DK2610351T3 (da) | 2007-12-05 | 2008-12-05 | Effektiv basebestemmelse i sekvenseringsreaktioner |
Country Status (3)
Country | Link |
---|---|
EP (2) | EP2565279B1 (da) |
CN (1) | CN103290106B (da) |
DK (2) | DK2565279T3 (da) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20170349893A1 (en) * | 2014-11-26 | 2017-12-07 | Bgi Shenzhen | Method and reagent for constructing nucleic acid double-linker single-strand cyclical library |
US10479991B2 (en) * | 2014-11-26 | 2019-11-19 | Mgi Tech Co., Ltd | Method and reagent for constructing nucleic acid double-linker single-strand cyclical library |
EP3245517B1 (en) | 2015-10-07 | 2018-09-19 | Selma Diagnostics ApS | Flow system and methods for digital counting |
JP6925051B2 (ja) | 2016-07-29 | 2021-08-25 | セルマ・ダイアグノスティクス・アンパルトセルスカブSelma Diagnostics Aps | デジタル計数のための方法の改良 |
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2008
- 2008-12-05 EP EP12164915.6A patent/EP2565279B1/en active Active
- 2008-12-05 EP EP12191212.5A patent/EP2610351B1/en active Active
- 2008-12-05 DK DK12164915.6T patent/DK2565279T3/da active
- 2008-12-05 CN CN201310051522.XA patent/CN103290106B/zh active Active
- 2008-12-05 DK DK12191212.5T patent/DK2610351T3/da active
Also Published As
Publication number | Publication date |
---|---|
CN103290106B (zh) | 2015-07-29 |
EP2565279A1 (en) | 2013-03-06 |
EP2610351A1 (en) | 2013-07-03 |
CN103290106A (zh) | 2013-09-11 |
DK2610351T3 (da) | 2015-09-28 |
EP2565279B1 (en) | 2014-12-03 |
EP2610351B1 (en) | 2015-07-08 |
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