CA2050276C - Reactions d'allongement d'amorce - Google Patents

Reactions d'allongement d'amorce Download PDF

Info

Publication number: CA2050276C
Authority: CA; Canada
Prior art keywords: dna; agent; dna polymerase; polymerase; solution
Prior art date: 1989-04-12
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.): Expired - Lifetime

Application number

CA002050276A

Other languages

English (en)

Other versions

CA2050276A1 (fr

Inventor

Stanley Tabor

Charles C. Richardson

Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Harvard University

Original Assignee

Harvard University

Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)

1989-04-12

Filing date

1990-04-10

Publication date

2003-03-11

1990-04-10 Application filed by Harvard University filed Critical Harvard University

1990-10-13 Publication of CA2050276A1 publication Critical patent/CA2050276A1/fr

2003-03-11 Application granted granted Critical

2003-03-11 Publication of CA2050276C publication Critical patent/CA2050276C/fr

2010-04-10 Anticipated expiration legal-status Critical

Status Expired - Lifetime legal-status Critical Current

Links

238000006243 chemical reaction Methods 0.000 title claims description 39
XPPKVPWEQAFLFU-UHFFFAOYSA-J diphosphate(4-) Chemical compound [O-]P([O-])(=O)OP([O-])([O-])=O XPPKVPWEQAFLFU-UHFFFAOYSA-J 0.000 claims abstract description 36
235000011180 diphosphates Nutrition 0.000 claims abstract description 36
239000011541 reaction mixture Substances 0.000 claims abstract description 13
239000003795 chemical substances by application Substances 0.000 claims description 53
239000013615 primer Substances 0.000 claims description 43
108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 40
102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 40
102000009609 Pyrophosphatases Human genes 0.000 claims description 32
108010009413 Pyrophosphatases Proteins 0.000 claims description 32
238000000034 method Methods 0.000 claims description 27
238000003752 polymerase chain reaction Methods 0.000 claims description 15
235000011178 triphosphate Nutrition 0.000 claims description 13
239000001226 triphosphate Substances 0.000 claims description 13
230000000694 effects Effects 0.000 claims description 10
238000001712 DNA sequencing Methods 0.000 claims description 9
239000000203 mixture Substances 0.000 claims description 8
102000004190 Enzymes Human genes 0.000 claims description 7
108090000790 Enzymes Proteins 0.000 claims description 7
108010006785 Taq Polymerase Proteins 0.000 claims description 6
UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 claims description 6
102100034343 Integrase Human genes 0.000 claims description 5
108010092799 RNA-directed DNA polymerase Proteins 0.000 claims description 5
238000000137 annealing Methods 0.000 claims description 4
239000003155 DNA primer Substances 0.000 claims description 3
238000006116 polymerization reaction Methods 0.000 claims description 3
108091034117 Oligonucleotide Proteins 0.000 abstract description 8
108020004414 DNA Proteins 0.000 description 22
239000000243 solution Substances 0.000 description 17
PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 12
FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
239000000872 buffer Substances 0.000 description 7
238000010438 heat treatment Methods 0.000 description 6
BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 5
229910052921 ammonium sulfate Inorganic materials 0.000 description 5
235000011130 ammonium sulphate Nutrition 0.000 description 5
239000011780 sodium chloride Substances 0.000 description 5
239000006228 supernatant Substances 0.000 description 5
125000002264 triphosphate group Chemical class [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 5
KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 4
108020004707 nucleic acids Proteins 0.000 description 4
102000039446 nucleic acids Human genes 0.000 description 4
150000007523 nucleic acids Chemical class 0.000 description 4
238000000746 purification Methods 0.000 description 4
102000053602 DNA Human genes 0.000 description 3
240000004808 Saccharomyces cerevisiae Species 0.000 description 3
239000002777 nucleoside Substances 0.000 description 3
239000000047 product Substances 0.000 description 3
230000002829 reductive effect Effects 0.000 description 3
102000009617 Inorganic Pyrophosphatase Human genes 0.000 description 2
108010009595 Inorganic Pyrophosphatase Proteins 0.000 description 2
TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
101710172880 Thioredoxin F-type, chloroplastic Proteins 0.000 description 2
239000007983 Tris buffer Substances 0.000 description 2
238000009825 accumulation Methods 0.000 description 2
238000003556 assay Methods 0.000 description 2
239000000499 gel Substances 0.000 description 2
230000000670 limiting effect Effects 0.000 description 2
-1 nucleoside triphosphates Chemical class 0.000 description 2
230000002441 reversible effect Effects 0.000 description 2
LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
WEEMDRWIKYCTQM-UHFFFAOYSA-N 2,6-dimethoxybenzenecarbothioamide Chemical compound COC1=CC=CC(OC)=C1C(N)=S WEEMDRWIKYCTQM-UHFFFAOYSA-N 0.000 description 1
QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
241000894006 Bacteria Species 0.000 description 1
108020004635 Complementary DNA Proteins 0.000 description 1
108020001019 DNA Primers Proteins 0.000 description 1
230000004543 DNA replication Effects 0.000 description 1
108010010803 Gelatin Proteins 0.000 description 1
229910019142 PO4 Inorganic materials 0.000 description 1
QTENRWWVYAAPBI-YZTFXSNBSA-N Streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@H]1[C@H](N=C(N)N)[C@@H](O)[C@H](N=C(N)N)[C@@H](O)[C@@H]1O QTENRWWVYAAPBI-YZTFXSNBSA-N 0.000 description 1
229930006000 Sucrose Natural products 0.000 description 1
CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
241000589596 Thermus Species 0.000 description 1
102000002933 Thioredoxin Human genes 0.000 description 1
240000001085 Trapa natans Species 0.000 description 1
238000003916 acid precipitation Methods 0.000 description 1
230000003321 amplification Effects 0.000 description 1
238000010804 cDNA synthesis Methods 0.000 description 1
230000015556 catabolic process Effects 0.000 description 1
238000005119 centrifugation Methods 0.000 description 1
239000007795 chemical reaction product Substances 0.000 description 1
239000002299 complementary DNA Substances 0.000 description 1
238000006731 degradation reaction Methods 0.000 description 1
230000001627 detrimental effect Effects 0.000 description 1
238000012631 diagnostic technique Methods 0.000 description 1
QPXWUAQRJLSJRT-UHFFFAOYSA-N diethoxyphosphinothioyl diethyl phosphate Chemical compound CCOP(=O)(OCC)OP(=S)(OCC)OCC QPXWUAQRJLSJRT-UHFFFAOYSA-N 0.000 description 1
239000012153 distilled water Substances 0.000 description 1
239000012634 fragment Substances 0.000 description 1
239000008273 gelatin Substances 0.000 description 1
229920000159 gelatin Polymers 0.000 description 1
235000019322 gelatine Nutrition 0.000 description 1
235000011852 gelatine desserts Nutrition 0.000 description 1
239000001963 growth medium Substances 0.000 description 1
230000007062 hydrolysis Effects 0.000 description 1
238000006460 hydrolysis reaction Methods 0.000 description 1
230000002401 inhibitory effect Effects 0.000 description 1
230000005764 inhibitory process Effects 0.000 description 1
239000007788 liquid Substances 0.000 description 1
229910001629 magnesium chloride Inorganic materials 0.000 description 1
238000004519 manufacturing process Methods 0.000 description 1
238000005259 measurement Methods 0.000 description 1
229910021645 metal ion Inorganic materials 0.000 description 1
238000002703 mutagenesis Methods 0.000 description 1
231100000350 mutagenesis Toxicity 0.000 description 1
238000003199 nucleic acid amplification method Methods 0.000 description 1
150000003833 nucleoside derivatives Chemical class 0.000 description 1
239000002773 nucleotide Substances 0.000 description 1
125000003729 nucleotide group Chemical group 0.000 description 1
239000008188 pellet Substances 0.000 description 1
239000012071 phase Substances 0.000 description 1
NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
239000010452 phosphate Substances 0.000 description 1
229940080469 phosphocellulose Drugs 0.000 description 1
229920002401 polyacrylamide Polymers 0.000 description 1
238000002360 preparation method Methods 0.000 description 1
230000002265 prevention Effects 0.000 description 1
230000005180 public health Effects 0.000 description 1
238000012163 sequencing technique Methods 0.000 description 1
239000007790 solid phase Substances 0.000 description 1
238000010561 standard procedure Methods 0.000 description 1
229960002385 streptomycin sulfate Drugs 0.000 description 1
239000005720 sucrose Substances 0.000 description 1
108060008226 thioredoxin Proteins 0.000 description 1
229940094937 thioredoxin Drugs 0.000 description 1
XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1

Classifications

- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
- 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
- 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
- C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction

Landscapes

Chemical & Material Sciences (AREA)
Life Sciences & Earth Sciences (AREA)
Organic Chemistry (AREA)
Health & Medical Sciences (AREA)
Engineering & Computer Science (AREA)
Zoology (AREA)
Wood Science & Technology (AREA)
Genetics & Genomics (AREA)
Bioinformatics & Cheminformatics (AREA)
Proteomics, Peptides & Aminoacids (AREA)
General Health & Medical Sciences (AREA)
Microbiology (AREA)
Molecular Biology (AREA)
Biotechnology (AREA)
General Engineering & Computer Science (AREA)
Biochemistry (AREA)
Analytical Chemistry (AREA)
Physics & Mathematics (AREA)
Immunology (AREA)
Biophysics (AREA)
Chemical Kinetics & Catalysis (AREA)
Medicinal Chemistry (AREA)
Biomedical Technology (AREA)
Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Enzymes And Modification Thereof (AREA)
Saccharide Compounds (AREA)

CA002050276A 1989-04-12 1990-04-10 Reactions d'allongement d'amorce Expired - Lifetime CA2050276C (fr)

Applications Claiming Priority (3)

Application Number	Priority Date	Filing Date	Title
US33675189A	1989-04-12	1989-04-12
US336,751		1989-04-12
PCT/US1990/001938 WO1990012111A1 (fr)	1989-04-12	1990-04-10	Reactions d'extension d'amorce ameliorees

Publications (2)

Publication Number	Publication Date
CA2050276A1 CA2050276A1 (fr)	1990-10-13
CA2050276C true CA2050276C (fr)	2003-03-11

Family

ID=23317491

Family Applications (1)

Application Number	Title	Priority Date	Filing Date
CA002050276A Expired - Lifetime CA2050276C (fr)	1989-04-12	1990-04-10	Reactions d'allongement d'amorce

Country Status (8)

Country	Link
EP (1)	EP0467953A4 (fr)
JP (1)	JP2997043B2 (fr)
KR (1)	KR920700294A (fr)
AU (1)	AU638246B2 (fr)
CA (1)	CA2050276C (fr)
HU (1)	HUT61054A (fr)
LT (1)	LTIP1519A (fr)
WO (1)	WO1990012111A1 (fr)

Families Citing this family (15)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
US5001050A (en) *	1989-03-24	1991-03-19	Consejo Superior Investigaciones Cientificas	PHφ29 DNA polymerase
US5198543A (en) *	1989-03-24	1993-03-30	Consejo Superior Investigaciones Cientificas	PHI29 DNA polymerase
FR2674254B1 (fr) *	1991-03-20	1995-10-06	Univ Reims Champagne Ardenne	Detection non radioactive de la presence d'un acide nucleique determine dans un echantillon biologique.
US5256555A (en)	1991-12-20	1993-10-26	Ambion, Inc.	Compositions and methods for increasing the yields of in vitro RNA transcription and other polynucleotide synthetic reactions
FI923911A7 (fi) *	1992-09-01	1994-03-02	Timo Korpela	DNA-molekyylien in vitro-synteesi
DE4336266A1 (de) *	1993-10-23	1995-04-27	Boehringer Mannheim Gmbh	Stabilisierte flüssige Mischungen für die Markierung von Nukleinsäuren
CA2222744C (fr) *	1995-05-31	2008-03-25	Amersham Life Science, Inc.	Adn-polymerases thermostables
US5665551A (en) *	1995-09-13	1997-09-09	Roche Molecular Systems, Inc.	Purified nucleic acid encoding a thermostable pyrophosphatase
DE19612779A1 (de) *	1996-03-29	1997-10-02	Boehringer Mannheim Gmbh	Verfahren zur spezifischen Vervielfältigung von langen Nukleinsäuren durch PCR
AU4652997A (en) *	1996-10-07	1998-05-05	Perkin-Elmer Corporation, The	Primer extension reaction utilizing a cosubstrate-enzyme pair for consuming pyrophosphate
US6291164B1 (en)	1996-11-22	2001-09-18	Invitrogen Corporation	Methods for preventing inhibition of nucleic acid synthesis by pyrophosphate
GB0110501D0 (en)	2001-04-30	2001-06-20	Secr Defence Brit	Amplification process
CA2540875A1 (fr) *	2003-09-30	2005-04-14	Perkinelmer Las, Inc.	Compositions et procedes de genotypage de polymorphismes d'un nucleotide simple
ES2994071T3 (en) *	2013-05-24	2025-01-16	Illumina Cambridge Ltd	Pyrophosphorolytic sequencing using nanopores
WO2022155282A1 (fr) *	2021-01-14	2022-07-21	Integrated Dna Technologies, Inc.	Procédés de production et de quantification de billes marquées par un identifiant moléculaire unique

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number	Priority date	Publication date	Assignee	Title
US4683202A (en) *	1985-03-28	1987-07-28	Cetus Corporation	Process for amplifying nucleic acid sequences
CA1338457C (fr) *	1986-08-22	1996-07-16	Henry A. Erlich	Enzyme thermostable purifiee
US4971903A (en) *	1988-03-25	1990-11-20	Edward Hyman	Pyrophosphate-based method and apparatus for sequencing nucleic acids

1990
- 1990-04-10 CA CA002050276A patent/CA2050276C/fr not_active Expired - Lifetime
- 1990-04-10 WO PCT/US1990/001938 patent/WO1990012111A1/fr not_active Ceased
- 1990-04-10 AU AU54382/90A patent/AU638246B2/en not_active Ceased
- 1990-04-10 EP EP19900906516 patent/EP0467953A4/en not_active Withdrawn
- 1990-04-10 JP JP2506210A patent/JP2997043B2/ja not_active Expired - Lifetime
- 1990-04-10 HU HU903562A patent/HUT61054A/hu unknown
- 1990-04-10 KR KR1019900702622A patent/KR920700294A/ko not_active Withdrawn
1993
- 1993-12-03 LT LTIP1519A patent/LTIP1519A/xx unknown

Also Published As

Publication number	Publication date
LTIP1519A (en)	1995-06-26
JPH04506002A (ja)	1992-10-22
WO1990012111A1 (fr)	1990-10-18
AU5438290A (en)	1990-11-05
CA2050276A1 (fr)	1990-10-13
KR920700294A (ko)	1992-02-19
HU903562D0 (en)	1992-03-30
HUT61054A (en)	1992-11-30
JP2997043B2 (ja)	2000-01-11
AU638246B2 (en)	1993-06-24
EP0467953A4 (en)	1992-06-17
EP0467953A1 (fr)	1992-01-29

Legal Events

Date	Code	Title
1997-04-10	EEER	Examination request
1998-12-02	FZDC	Correction of dead application (reinstatement)
2010-04-10	MKEX	Expiry

Publication	Publication Date	Title
US5498523A (en)	1996-03-12	DNA sequencing with pyrophosphatase
EP0527728B1 (fr)	1997-07-09	Reactions de synthese d'adn (in vitro) utilisant la polymerase de phi 29 adn modifiee, et un fragment d'adn codant ladite polymerase
AU773536B2 (en)	2004-05-27	Method for amplifying nucleic acid sequence
EP0386858B1 (fr)	1994-04-13	T7 ADN polymérase
US5773257A (en)	1998-06-30	Method for producing primed nucleic acid templates
JP3421664B2 (ja)	2003-06-30	ヌクレオチド塩基の同定方法
US4921794A (en)	1990-05-01	T7 DNA polymerase
US5648211A (en)	1997-07-15	Strand displacement amplification using thermophilic enzymes
US5576204A (en)	1996-11-19	φ29 DNA polymerase
US4795699A (en)	1989-01-03	T7 DNA polymerase
US4994372A (en)	1991-02-19	DNA sequencing
US7501237B2 (en)	2009-03-10	Polymerases for analyzing or typing polymorphic nucleic acid fragments and uses thereof
CA2421492C (fr)	2010-05-04	Methode de sequencage de l'adn
EP0655506A1 (fr)	1995-05-31	Polymérase d'ADN ayant les nucléotiden modifiées pour le site d'adhésion de l'ADN
AU638246B2 (en)	1993-06-24	Improved primer extension reactions
US5605824A (en)	1997-02-25	Composition for hybridizing nucleic acids using single-stranded nucleic acid binding protein
US5173411A (en)	1992-12-22	Method for determining the nucleotide base sequence of a DNA molecule
US20030186312A1 (en)	2003-10-02	Method for synthesizing DNA
WO1994005797A1 (fr)	1994-03-17	Synthese in vitro de molecules d'adn
WO1991006679A1 (fr)	1991-05-16	Procede ameliore d'hybridation des acides nucleiques utilisant une proteine monocatenaire liante de l'acide nucleique
US5516633A (en)	1996-05-14	DNA sequencing with a T7-type gene 6 exonuclease
US6048696A (en)	2000-04-11	Method of identifying nucleic acid molecules
Tabor et al.	1995	DNA sequencing with pyrophosphatase
KR100484124B1 (ko)	2005-09-08	변형된 뉴클레오티드 결합부위를 갖는 dna 서열화용 dna 폴리머라제
Scherzinger et al.	1974	Initiation of the Replication of Single‐Stranded DNA by Concerted Action of Phage T7 RNA and DNA Polymerases