CA2167840A1 - Amplifications lineaires successives d'acides nucleiques - Google Patents
Amplifications lineaires successives d'acides nucleiquesInfo
- Publication number
- CA2167840A1 CA2167840A1 CA002167840A CA2167840A CA2167840A1 CA 2167840 A1 CA2167840 A1 CA 2167840A1 CA 002167840 A CA002167840 A CA 002167840A CA 2167840 A CA2167840 A CA 2167840A CA 2167840 A1 CA2167840 A1 CA 2167840A1
- Authority
- CA
- Canada
- Prior art keywords
- primers
- primer extension
- primer
- nucleic acid
- process according
- Prior art date
- 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.)
- Abandoned
Links
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 82
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 64
- 230000003321 amplification Effects 0.000 title claims abstract description 62
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 48
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 48
- 238000006243 chemical reaction Methods 0.000 claims abstract description 105
- 238000000034 method Methods 0.000 claims abstract description 88
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 35
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 13
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 12
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 12
- 238000001668 nucleic acid synthesis Methods 0.000 claims abstract description 7
- 230000000295 complement effect Effects 0.000 claims description 22
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 claims description 17
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 claims description 17
- DNIAPMSPPWPWGF-VKHMYHEASA-N (+)-propylene glycol Chemical compound C[C@H](O)CO DNIAPMSPPWPWGF-VKHMYHEASA-N 0.000 claims description 11
- 208000026350 Inborn Genetic disease Diseases 0.000 claims description 11
- 208000016361 genetic disease Diseases 0.000 claims description 11
- 229940035437 1,3-propanediol Drugs 0.000 claims description 9
- YPFDHNVEDLHUCE-UHFFFAOYSA-N 1,3-propanediol Substances OCCCO YPFDHNVEDLHUCE-UHFFFAOYSA-N 0.000 claims description 9
- 229920000166 polytrimethylene carbonate Polymers 0.000 claims description 9
- 239000011541 reaction mixture Substances 0.000 claims description 9
- 239000005547 deoxyribonucleotide Substances 0.000 claims description 5
- 125000002637 deoxyribonucleotide group Chemical group 0.000 claims description 5
- 108020004711 Nucleic Acid Probes Proteins 0.000 claims description 3
- 239000002853 nucleic acid probe Substances 0.000 claims description 3
- 208000007056 sickle cell anemia Diseases 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- NSMOSDAEGJTOIQ-CRCLSJGQSA-N (2r,3s)-2-(hydroxymethyl)oxolan-3-ol Chemical compound OC[C@H]1OCC[C@@H]1O NSMOSDAEGJTOIQ-CRCLSJGQSA-N 0.000 claims 7
- 108091028664 Ribonucleotide Proteins 0.000 claims 3
- 239000002336 ribonucleotide Substances 0.000 claims 3
- 125000002652 ribonucleotide group Chemical group 0.000 claims 3
- 230000005856 abnormality Effects 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000009825 accumulation Methods 0.000 abstract description 11
- 238000001514 detection method Methods 0.000 abstract description 7
- 238000009877 rendering Methods 0.000 abstract 1
- 239000013615 primer Substances 0.000 description 262
- 108020004414 DNA Proteins 0.000 description 83
- 239000000872 buffer Substances 0.000 description 17
- 239000012528 membrane Substances 0.000 description 15
- 239000000523 sample Substances 0.000 description 14
- 230000006820 DNA synthesis Effects 0.000 description 13
- 125000003729 nucleotide group Chemical group 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 11
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 10
- 238000009396 hybridization Methods 0.000 description 10
- 239000002773 nucleotide Substances 0.000 description 10
- 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 9
- 108700028369 Alleles Proteins 0.000 description 9
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- 102000053602 DNA Human genes 0.000 description 8
- ULWHHBHJGPPBCO-UHFFFAOYSA-N propane-1,1-diol Chemical group CCC(O)O ULWHHBHJGPPBCO-UHFFFAOYSA-N 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
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- OSBLTNPMIGYQGY-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid;boric acid Chemical compound OB(O)O.OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O OSBLTNPMIGYQGY-UHFFFAOYSA-N 0.000 description 6
- 239000003155 DNA primer Substances 0.000 description 6
- 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 6
- 102000004190 Enzymes Human genes 0.000 description 6
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- 101000914338 Mus musculus Carcinoembryonic antigen-related cell adhesion molecule 2 Proteins 0.000 description 6
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- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 239000012634 fragment Substances 0.000 description 6
- 238000005382 thermal cycling Methods 0.000 description 6
- 229920001917 Ficoll Polymers 0.000 description 5
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 5
- 208000037065 Subacute sclerosing leukoencephalitis Diseases 0.000 description 5
- 206010042297 Subacute sclerosing panencephalitis Diseases 0.000 description 5
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 5
- 238000004925 denaturation Methods 0.000 description 5
- 230000036425 denaturation Effects 0.000 description 5
- 229910001629 magnesium chloride Inorganic materials 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 108091093088 Amplicon Proteins 0.000 description 4
- 230000004544 DNA amplification Effects 0.000 description 4
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- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 4
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 4
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 4
- 239000012160 loading buffer Substances 0.000 description 4
- 229920001778 nylon Polymers 0.000 description 4
- 230000037452 priming Effects 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 3
- 108010017826 DNA Polymerase I Proteins 0.000 description 3
- 102000004594 DNA Polymerase I Human genes 0.000 description 3
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- RZCIEJXAILMSQK-JXOAFFINSA-N TTP Chemical compound O=C1NC(=O)C(C)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 RZCIEJXAILMSQK-JXOAFFINSA-N 0.000 description 3
- 108010006785 Taq Polymerase Proteins 0.000 description 3
- 108010058966 bacteriophage T7 induced DNA polymerase Proteins 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
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- 239000011780 sodium chloride Substances 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- VLEIUWBSEKKKFX-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O VLEIUWBSEKKKFX-UHFFFAOYSA-N 0.000 description 2
- 208000035657 Abasia Diseases 0.000 description 2
- 235000014676 Phragmites communis Nutrition 0.000 description 2
- 108020004682 Single-Stranded DNA Proteins 0.000 description 2
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 2
- 238000007844 allele-specific PCR Methods 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001186 cumulative effect Effects 0.000 description 2
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- 238000013461 design Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 2
- 229960005542 ethidium bromide Drugs 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000006116 polymerization reaction Methods 0.000 description 2
- 230000003362 replicative effect Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- JEPVUMTVFPQKQE-AAKCMJRZSA-N 2-[(1s,2s,3r,4s)-1,2,3,4,5-pentahydroxypentyl]-1,3-thiazolidine-4-carboxylic acid Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C1NC(C(O)=O)CS1 JEPVUMTVFPQKQE-AAKCMJRZSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102100024533 Carcinoembryonic antigen-related cell adhesion molecule 1 Human genes 0.000 description 1
- 101710190843 Carcinoembryonic antigen-related cell adhesion molecule 1 Proteins 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 240000006497 Dianthus caryophyllus Species 0.000 description 1
- 235000009355 Dianthus caryophyllus Nutrition 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000004327 boric acid Substances 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000009390 chemical decontamination Methods 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 1
- 239000005549 deoxyribonucleoside Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000011143 downstream manufacturing Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000002474 experimental method Methods 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
- 238000012215 gene cloning Methods 0.000 description 1
- 108060003196 globin Proteins 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- LCNBIHVSOPXFMR-UHFFFAOYSA-N n'-(3-aminopropyl)butane-1,4-diamine;hydron;trichloride Chemical compound Cl.Cl.Cl.NCCCCNCCCN LCNBIHVSOPXFMR-UHFFFAOYSA-N 0.000 description 1
- 125000001624 naphthyl group Chemical group 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000002987 primer (paints) Substances 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 239000002342 ribonucleoside Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- NLIVDORGVGAOOJ-MAHBNPEESA-M xylene cyanol Chemical compound [Na+].C1=C(C)C(NCC)=CC=C1C(\C=1C(=CC(OS([O-])=O)=CC=1)OS([O-])=O)=C\1C=C(C)\C(=[NH+]/CC)\C=C/1 NLIVDORGVGAOOJ-MAHBNPEESA-M 0.000 description 1
Classifications
-
- 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)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Saccharide Compounds (AREA)
Abstract
On effectue la synthèse extensive ("amplification") d'une séquence d'acide nucléique à étudier au moyen d'une série enchaînée de réactions d'extension d'amorces à plusieurs cycles. Les amorces utilisées dans chacune des réactions d'extension d'amorce du procédé contiennent des éléments non réplicables qui stoppent la synthèse d'acide nucléique et empêchent par conséquent les molécules synthétisées de servir de matrice dans les cycles subséquents. Les molécules synthétisées s'accumulent de manière mathématiquement linéaire pendant l'extension de l'amorce et confèrent au procédé une relative insensibilité par rapport aux acides nucléiques contaminants. Plusieurs ensembles d'amorces sont utilisés afin d'assurer l'accumulation d'un grand nombre de copies de la séquence d'acide nucléique à étudier. L'invention concerne également la détection d'une séquence d'acide nucléique à étudier ainsi que des trousses de réactifs permettant d'effectuer une telle réaction.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US9544293A | 1993-07-23 | 1993-07-23 | |
| US08/095,442 | 1993-07-23 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2167840A1 true CA2167840A1 (fr) | 1995-02-02 |
Family
ID=22252059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002167840A Abandoned CA2167840A1 (fr) | 1993-07-23 | 1994-07-25 | Amplifications lineaires successives d'acides nucleiques |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0710296A4 (fr) |
| JP (1) | JPH09500539A (fr) |
| CN (1) | CN1127533A (fr) |
| CA (1) | CA2167840A1 (fr) |
| WO (1) | WO1995003432A1 (fr) |
Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6335184B1 (en) | 1993-07-23 | 2002-01-01 | Bio-Rad Laboratories, Inc. | Linked linear amplification of nucleic acids |
| CN1117871C (zh) * | 1995-06-07 | 2003-08-13 | 伯乐实验有限公司 | 核酸的衔接线性扩增 |
| US5854033A (en) | 1995-11-21 | 1998-12-29 | Yale University | Rolling circle replication reporter systems |
| ATE199572T1 (de) | 1995-11-21 | 2001-03-15 | Univ Yale | Unimolekulare segmentamplifikation und bestimmung |
| WO2000004193A1 (fr) | 1998-07-20 | 2000-01-27 | Yale University | Procede pour detecter des acides nucleiques au moyen de ligature a mediation par cibles d'amorces bipartites |
| WO2000015779A2 (fr) | 1998-09-15 | 2000-03-23 | Yale University | Clonage moleculaire a amplification selon le modele du cercle roulant |
| JP2002525078A (ja) | 1998-09-15 | 2002-08-13 | イェール ユニバーシティ | 人工的長末端反復配列 |
| US7205129B1 (en) * | 2000-02-28 | 2007-04-17 | Qiagen Gmbh | Method for reducing artifacts in nucleic acid amplification |
| ATE374835T1 (de) | 2000-06-30 | 2007-10-15 | Qiagen Gmbh | SIGNALAMPLIFIKATION MIT ßLOLLIPOPß- HYDRIDISIERUNGSPROBEN |
| US6573051B2 (en) | 2001-03-09 | 2003-06-03 | Molecular Staging, Inc. | Open circle probes with intramolecular stem structures |
| US7553619B2 (en) | 2002-02-08 | 2009-06-30 | Qiagen Gmbh | Detection method using dissociated rolling circle amplification |
| US9487823B2 (en) | 2002-12-20 | 2016-11-08 | Qiagen Gmbh | Nucleic acid amplification |
| US6977153B2 (en) | 2002-12-31 | 2005-12-20 | Qiagen Gmbh | Rolling circle amplification of RNA |
| EP1762627A1 (fr) | 2005-09-09 | 2007-03-14 | Qiagen GmbH | Procédé pour l'activation d'acides nucléiques pour effectuer une réaction d'une polymérase |
| EP2377929A1 (fr) | 2005-10-06 | 2011-10-19 | Lucigen Corporation | Polymérases virales thermostables et procédés d'utilisation |
| CA2923083C (fr) * | 2013-09-13 | 2018-07-17 | F. Hoffmann-La Roche Ag | Application de molecules oligo-dt pour eviter la production de produits de pcr de masse moleculaire elevee induite par un transporteur de polya |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4683195A (en) * | 1986-01-30 | 1987-07-28 | Cetus Corporation | Process for amplifying, detecting, and/or-cloning nucleic acid sequences |
| IE61148B1 (en) * | 1988-03-10 | 1994-10-05 | Ici Plc | Method of detecting nucleotide sequences |
| GB8920097D0 (en) * | 1989-09-06 | 1989-10-18 | Ici Plc | Amplification processes |
-
1994
- 1994-07-25 CA CA002167840A patent/CA2167840A1/fr not_active Abandoned
- 1994-07-25 EP EP94924010A patent/EP0710296A4/fr not_active Withdrawn
- 1994-07-25 WO PCT/US1994/008390 patent/WO1995003432A1/fr not_active Ceased
- 1994-07-25 JP JP7505384A patent/JPH09500539A/ja active Pending
- 1994-07-25 CN CN94192853A patent/CN1127533A/zh active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| EP0710296A1 (fr) | 1996-05-08 |
| JPH09500539A (ja) | 1997-01-21 |
| EP0710296A4 (fr) | 1999-05-26 |
| WO1995003432A1 (fr) | 1995-02-02 |
| CN1127533A (zh) | 1996-07-24 |
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| Date | Code | Title | Description |
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| EEER | Examination request | ||
| FZDE | Dead |