JP5191041B2 - 急速ワンステップｒｔ−ｐｃｒ - Google Patents
急速ワンステップｒｔ−ｐｃｒ Download PDFInfo
- Publication number
- JP5191041B2 JP5191041B2 JP2008094907A JP2008094907A JP5191041B2 JP 5191041 B2 JP5191041 B2 JP 5191041B2 JP 2008094907 A JP2008094907 A JP 2008094907A JP 2008094907 A JP2008094907 A JP 2008094907A JP 5191041 B2 JP5191041 B2 JP 5191041B2
- Authority
- JP
- Japan
- Prior art keywords
- pcr
- temperature
- polymerase
- seconds
- reverse transcriptase
- 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.)
- Active
Links
- 238000003757 reverse transcription PCR Methods 0.000 title claims description 43
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 claims description 48
- 102100034343 Integrase Human genes 0.000 claims description 47
- 238000006243 chemical reaction Methods 0.000 claims description 42
- 238000000034 method Methods 0.000 claims description 37
- 230000000694 effects Effects 0.000 claims description 22
- 239000002299 complementary DNA Substances 0.000 claims description 17
- 230000001419 dependent effect Effects 0.000 claims description 14
- 239000000203 mixture Substances 0.000 claims description 13
- 238000005382 thermal cycling Methods 0.000 claims description 11
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 239000011541 reaction mixture Substances 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 2
- 239000000523 sample Substances 0.000 description 42
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 31
- 238000011533 pre-incubation Methods 0.000 description 25
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 17
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 17
- 238000009396 hybridization Methods 0.000 description 14
- 108020004414 DNA Proteins 0.000 description 13
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 238000002866 fluorescence resonance energy transfer Methods 0.000 description 11
- 238000010839 reverse transcription Methods 0.000 description 11
- 230000003321 amplification Effects 0.000 description 10
- 238000003199 nucleic acid amplification method Methods 0.000 description 10
- 238000011534 incubation Methods 0.000 description 8
- 108020004999 messenger RNA Proteins 0.000 description 7
- 238000004925 denaturation Methods 0.000 description 6
- 230000036425 denaturation Effects 0.000 description 6
- 230000014509 gene expression Effects 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 150000007523 nucleic acids Chemical group 0.000 description 5
- 241000713838 Avian myeloblastosis virus Species 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 4
- 108010006785 Taq Polymerase Proteins 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000007062 hydrolysis Effects 0.000 description 4
- 238000006460 hydrolysis reaction Methods 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 238000003753 real-time PCR Methods 0.000 description 4
- 102100028161 ATP-binding cassette sub-family C member 2 Human genes 0.000 description 3
- 108010066419 Multidrug Resistance-Associated Protein 2 Proteins 0.000 description 3
- 241000589596 Thermus Species 0.000 description 3
- 108010085671 Thermus thermophilus DNA polymerase Proteins 0.000 description 3
- 108010020713 Tth polymerase Proteins 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 230000007704 transition Effects 0.000 description 3
- 241000620141 Carboxydothermus Species 0.000 description 2
- 108060002716 Exonuclease Proteins 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- 241000713869 Moloney murine leukemia virus Species 0.000 description 2
- 108010010677 Phosphodiesterase I Proteins 0.000 description 2
- 241000589499 Thermus thermophilus Species 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 238000010804 cDNA synthesis Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
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- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 102000013165 exonuclease Human genes 0.000 description 2
- 239000000833 heterodimer Substances 0.000 description 2
- 229910001437 manganese ion Inorganic materials 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 208000030507 AIDS Diseases 0.000 description 1
- 108091093088 Amplicon Proteins 0.000 description 1
- 101000909256 Caldicellulosiruptor bescii (strain ATCC BAA-1888 / DSM 6725 / Z-1320) DNA polymerase I Proteins 0.000 description 1
- 241000620137 Carboxydothermus hydrogenoformans Species 0.000 description 1
- 241001112695 Clostridiales Species 0.000 description 1
- 230000004568 DNA-binding Effects 0.000 description 1
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 description 1
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 1
- 241000204652 Thermotoga Species 0.000 description 1
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- 241001493065 dsRNA viruses Species 0.000 description 1
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- 238000010195 expression analysis Methods 0.000 description 1
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- 230000006872 improvement Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000009830 intercalation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 229910001425 magnesium ion Inorganic materials 0.000 description 1
- 229940071125 manganese acetate Drugs 0.000 description 1
- UOGMEBQRZBEZQT-UHFFFAOYSA-L manganese(2+);diacetate Chemical compound [Mn+2].CC([O-])=O.CC([O-])=O UOGMEBQRZBEZQT-UHFFFAOYSA-L 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000011880 melting curve analysis Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 239000002751 oligonucleotide probe Substances 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 230000001915 proofreading effect Effects 0.000 description 1
- 230000020978 protein processing Effects 0.000 description 1
- 230000007026 protein scission Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- ABZLKHKQJHEPAX-UHFFFAOYSA-N tetramethylrhodamine Chemical compound C=12C=CC(N(C)C)=CC2=[O+]C2=CC(N(C)C)=CC=C2C=1C1=CC=CC=C1C([O-])=O ABZLKHKQJHEPAX-UHFFFAOYSA-N 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000005758 transcription activity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000011144 upstream manufacturing Methods 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
- C12Q1/6846—Common amplification features
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- 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)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP07007173 | 2007-04-05 | ||
| EP07007173.3 | 2007-04-05 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| JP2008253263A JP2008253263A (ja) | 2008-10-23 |
| JP2008253263A5 JP2008253263A5 (enExample) | 2010-04-08 |
| JP5191041B2 true JP5191041B2 (ja) | 2013-04-24 |
Family
ID=38453943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2008094907A Active JP5191041B2 (ja) | 2007-04-05 | 2008-04-01 | 急速ワンステップｒｔ−ｐｃｒ |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US8119353B2 (enExample) |
| EP (1) | EP1978109B1 (enExample) |
| JP (1) | JP5191041B2 (enExample) |
| ES (1) | ES2398984T3 (enExample) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022138887A1 (ja) | 2020-12-25 | 2022-06-30 | 株式会社ニッポンジーン | 改変型ｄｎａポリメラーゼ |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9175332B2 (en) * | 2010-07-29 | 2015-11-03 | Roche Molecular Systems, Inc. | Generic PCR |
| ES2533732T3 (es) | 2010-10-04 | 2015-04-14 | F. Hoffmann-La Roche Ag | Método para lisis celular en un tampón de reacción de RT-PCR |
| WO2013101290A1 (en) * | 2011-12-29 | 2013-07-04 | Qiagen | Methods of using telomeres as markers for aging |
| PT2855692T (pt) | 2012-05-24 | 2019-10-25 | Univ Utah Res Found | Pcr extrema. |
| CN108367287B (zh) * | 2015-11-05 | 2021-04-02 | 犹他州大学研究基金会 | 极端逆转录pcr |
| JP2018023360A (ja) * | 2016-07-29 | 2018-02-15 | セイコーエプソン株式会社 | 核酸増幅反応方法、試薬、および試薬の使用方法 |
| GB201812192D0 (en) | 2018-07-26 | 2018-09-12 | Ttp Plc | Variable temperature reactor, heater and control circuit for the same |
| US20220064707A1 (en) * | 2019-01-17 | 2022-03-03 | Northwestern University | Rapid reverse transcription quantitative polymerase chain reaction |
Family Cites Families (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5105599A (en) * | 1989-02-24 | 1992-04-21 | Highland Supply Corporation | Means for securing a decorative cover about a flower pot |
| US5455170A (en) | 1986-08-22 | 1995-10-03 | Hoffmann-La Roche Inc. | Mutated thermostable nucleic acid polymerase enzyme from Thermus species Z05 |
| US5374553A (en) | 1986-08-22 | 1994-12-20 | Hoffmann-La Roche Inc. | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
| US5561058A (en) | 1986-08-22 | 1996-10-01 | Hoffmann-La Roche Inc. | Methods for coupled high temperatures reverse transcription and polymerase chain reactions |
| US5118801A (en) | 1988-09-30 | 1992-06-02 | The Public Health Research Institute | Nucleic acid process containing improved molecular switch |
| AU656315B2 (en) | 1989-12-22 | 1995-02-02 | F. Hoffmann-La Roche Ag | pigh temperature reverse transcriptases |
| US5210015A (en) | 1990-08-06 | 1993-05-11 | Hoffman-La Roche Inc. | Homogeneous assay system using the nuclease activity of a nucleic acid polymerase |
| CA2124581A1 (en) | 1992-09-29 | 1994-04-14 | John M. Vierling | One step pcr for detection of rare rna and dna |
| US5538848A (en) * | 1994-11-16 | 1996-07-23 | Applied Biosystems Division, Perkin-Elmer Corp. | Method for detecting nucleic acid amplification using self-quenching fluorescence probe |
| US5876924A (en) * | 1994-06-22 | 1999-03-02 | Mount Sinai School Of Medicine | Nucleic acid amplification method hybridization signal amplification method (HSAM) |
| CA2591550C (en) | 1996-06-04 | 2009-04-28 | University Of Utah Research Foundation | Monitoring hybridization during pcr |
| EP2298931B2 (en) | 1996-06-04 | 2020-06-24 | University of Utah Research Foundation | Apparatus for performing PCR and monitoring the reaction in real time during temperature cycling |
| US7179590B2 (en) | 2000-04-18 | 2007-02-20 | Roche Molecular Systems, Inc | High temperature reverse transcription using mutant DNA polymerases |
| CN100523216C (zh) * | 2001-03-02 | 2009-08-05 | 匹兹堡大学 | Pcr方法 |
| US20030108898A1 (en) * | 2001-07-03 | 2003-06-12 | Paul Choppa | Method for detection, quantitation and breakpoint cluster region determination of fusion transcripts |
| AU2004205774B2 (en) * | 2003-01-17 | 2006-12-14 | The Chinese University Of Hong Kong | Circulating mRNA as diagnostic markers for pregnancy-related disorders |
-
2008
- 2008-04-01 JP JP2008094907A patent/JP5191041B2/ja active Active
- 2008-04-03 EP EP08006778A patent/EP1978109B1/en active Active
- 2008-04-03 ES ES08006778T patent/ES2398984T3/es active Active
- 2008-04-03 US US12/061,700 patent/US8119353B2/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022138887A1 (ja) | 2020-12-25 | 2022-06-30 | 株式会社ニッポンジーン | 改変型ｄｎａポリメラーゼ |
Also Published As
| Publication number | Publication date |
|---|---|
| US20080248535A1 (en) | 2008-10-09 |
| EP1978109B1 (en) | 2012-12-05 |
| EP1978109A1 (en) | 2008-10-08 |
| ES2398984T3 (es) | 2013-03-25 |
| US8119353B2 (en) | 2012-02-21 |
| JP2008253263A (ja) | 2008-10-23 |
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