CA2211367A1 - Improved method for reverse transcription - Google Patents
Improved method for reverse transcriptionInfo
- Publication number
- CA2211367A1 CA2211367A1 CA002211367A CA2211367A CA2211367A1 CA 2211367 A1 CA2211367 A1 CA 2211367A1 CA 002211367 A CA002211367 A CA 002211367A CA 2211367 A CA2211367 A CA 2211367A CA 2211367 A1 CA2211367 A1 CA 2211367A1
- Authority
- CA
- Canada
- Prior art keywords
- mrna
- reverse transcription
- reverse transcriptase
- improved method
- metal ions
- 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.)
- Granted
Links
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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1096—Processes for the isolation, preparation or purification of DNA or RNA cDNA Synthesis; Subtracted cDNA library construction, e.g. RT, RT-PCR
-
- 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
Abstract
A method for preparing a cDNA from a mRNA using a reverse transcriptase wherein reverse transcription is performed at a temperature at which the mRNA does not take a secondary structure, for example, at a temperature of 45°C or more. The method is performed, for example, using a heat-labile reverse transcriptase in the presence of a substance exhibiting chaperone function having chaperone function such as saccharides. The method is performed, for example, in the presence of metal ions necessary for activation of the reverse transcriptase and a chelating agent for the metal ions such as a deoxynucleotide triphosphate. The method is capable of reverse transcription over the full length of mRNA template even if the mRNA is a long chain mRNA and, as a result, producing a full length cDNA.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA2512701A CA2512701C (en) | 1996-07-25 | 1997-07-24 | Improved method for reverse transcription |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP19632996 | 1996-07-25 | ||
JP19633196A JP3709490B2 (en) | 1996-07-25 | 1996-07-25 | Method for improving the thermal stability of RNA |
JP196329/1996 | 1996-07-25 | ||
JP196331/1996 | 1996-07-25 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA2512701A Division CA2512701C (en) | 1996-07-25 | 1997-07-24 | Improved method for reverse transcription |
Publications (2)
Publication Number | Publication Date |
---|---|
CA2211367A1 true CA2211367A1 (en) | 1998-01-25 |
CA2211367C CA2211367C (en) | 2008-04-22 |
Family
ID=26509679
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002211367A Expired - Fee Related CA2211367C (en) | 1996-07-25 | 1997-07-24 | Improved method for reverse transcription |
Country Status (4)
Country | Link |
---|---|
US (3) | US6013488A (en) |
EP (2) | EP1243661B1 (en) |
CA (1) | CA2211367C (en) |
DE (2) | DE69734913T2 (en) |
Families Citing this family (29)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CA2287542A1 (en) | 1997-04-22 | 1998-10-29 | Life Technologies, Inc. | Methods for the production of aslv reverse transcriptases composed of multiple subunits |
AU8162598A (en) * | 1997-06-25 | 1999-01-04 | Life Technologies, Inc. | Improved method for isolating and recovering target dna or rna molecules having a desired nucleotide sequence |
US6787305B1 (en) | 1998-03-13 | 2004-09-07 | Invitrogen Corporation | Compositions and methods for enhanced synthesis of nucleic acid molecules |
EP1129064B1 (en) | 1998-11-12 | 2008-01-09 | Invitrogen Corporation | Transfection reagents |
US7074556B2 (en) | 1999-03-02 | 2006-07-11 | Invitrogen Corporation | cDNA synthesis improvements |
WO2000055307A2 (en) * | 1999-03-12 | 2000-09-21 | Mcgill University | A method for increasing the processivity of a dna- or rna-dependent polymerase and compositions therefor |
US6271004B1 (en) * | 1999-06-25 | 2001-08-07 | Display Systems Biotech A/S | Method for improved reverse transcription at high temperatures |
US6835537B1 (en) | 2000-09-29 | 2004-12-28 | Myriad Genetics, Inc. | Method for equalizing band intensities on sequencing gels |
DE10105208A1 (en) * | 2001-02-06 | 2002-08-14 | Ihf Inst Fuer Hormon Und Fortp | Enzymatic production of cDNA by reverse transcription, in the presence of betaine, provides a high yield of full-length molecules from large RNA templates |
ES2188357B1 (en) * | 2001-03-12 | 2005-01-01 | BIOTOOLS BIOTECHNOLOGICAL & MEDICAL LABORATORIES, S.A. | METHOD FOR AMPLIFICATION BY TRANSCRIPTION IN REVERSE COUPLED TO A REACTION OF NESTED OR SEMI-NESTED PCR IN ONE STEP, OF A DIANA RNA, AND ITS APPLICATIONS IN THE IDENTIFICATION OF ENTEROVIRUS AND IN THE DISCRIMINATION BETWEEN POLIOVIRUS AND NON-POLIO. |
US20050250100A1 (en) * | 2002-06-12 | 2005-11-10 | Yoshihide Hayashizaki | Method of utilizing the 5'end of transcribed nucleic acid regions for cloning and analysis |
US20040152072A1 (en) * | 2002-07-30 | 2004-08-05 | Invitrogen Corporation | Reverse transcription |
US20060084799A1 (en) * | 2003-09-24 | 2006-04-20 | Williams Lewis T | Human cDNA clones comprising polynucleotides encoding polypeptides and methods of their use |
WO2005035569A2 (en) * | 2003-10-10 | 2005-04-21 | Five Prime Therapeutics, Inc. | Kiaa0779, splice variants thereof, and methods of their use |
US20080176209A1 (en) * | 2004-04-08 | 2008-07-24 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
CN101035620B (en) | 2004-04-08 | 2012-01-04 | 生物马特里卡公司 | Integration of sample storage and sample management for life science |
US20060099567A1 (en) * | 2004-04-08 | 2006-05-11 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
US7993853B2 (en) | 2005-05-06 | 2011-08-09 | Gen-Probe Incorporated | Methods of nucleic acid target capture |
EP2405024A1 (en) | 2005-07-15 | 2012-01-11 | Life Technologies Corporation | Hot start reverse transcription by primer design |
US9845489B2 (en) | 2010-07-26 | 2017-12-19 | Biomatrica, Inc. | Compositions for stabilizing DNA, RNA and proteins in saliva and other biological samples during shipping and storage at ambient temperatures |
WO2012018638A2 (en) | 2010-07-26 | 2012-02-09 | Biomatrica, Inc. | Compositions for stabilizing dna, rna and proteins in blood and other biological samples during shipping and storage at ambient temperatures |
US9458484B2 (en) | 2010-10-22 | 2016-10-04 | Bio-Rad Laboratories, Inc. | Reverse transcriptase mixtures with improved storage stability |
FR2984357B1 (en) * | 2011-12-16 | 2016-11-18 | Biomerieux Sa | PROCESS FOR TRANSCRIPTIONAL AMPLIFICATION OF NUCLEIC ACIDS COVERING STEPS OF DIFFERENT TEMPERATURES |
US9725703B2 (en) | 2012-12-20 | 2017-08-08 | Biomatrica, Inc. | Formulations and methods for stabilizing PCR reagents |
WO2015112767A2 (en) | 2014-01-22 | 2015-07-30 | Life Technologies Corporation | Novel reverse transcriptases for use in high temperature nucleic acid synthesis |
WO2015191632A1 (en) | 2014-06-10 | 2015-12-17 | Biomatrica, Inc. | Stabilization of thrombocytes at ambient temperatures |
EP3169310A1 (en) | 2014-07-15 | 2017-05-24 | Life Technologies Corporation | Compositions with lipid aggregates and methods for efficient delivery of molecules to cells |
WO2017100212A1 (en) | 2015-12-08 | 2017-06-15 | Biomatrica, Inc. | Reduction of erythrocyte sedimentation rate |
US11312991B2 (en) | 2016-01-27 | 2022-04-26 | Kabushiki Kaisha Dnaform | Method for decoding base sequence of nucleic acid corresponding to end region of RNA and method for analyzing DNA element |
Family Cites Families (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0035204B2 (en) * | 1980-03-05 | 1991-05-22 | Miles Inc. | Pasteurized therapeutically active protein compositions |
US4409200A (en) * | 1980-04-28 | 1983-10-11 | Research Corporation | Reverse transcriptase from human milk, method for its purification, and its use in the detection of breast cancer |
US5017492A (en) * | 1986-02-27 | 1991-05-21 | Life Technologies, Inc. | Reverse transcriptase and method for its production |
US5310652A (en) * | 1986-08-22 | 1994-05-10 | Hoffman-La Roche Inc. | Reverse transcription with thermostable DNA polymerase-high temperature reverse transcription |
US5407800A (en) * | 1986-08-22 | 1995-04-18 | Hoffmann-La Roche Inc. | Reverse transcription with Thermus thermophilus polymerase |
US5374553A (en) * | 1986-08-22 | 1994-12-20 | Hoffmann-La Roche Inc. | DNA encoding a thermostable nucleic acid polymerase enzyme from thermotoga maritima |
EP0506889B1 (en) * | 1989-12-22 | 1997-04-02 | F. Hoffmann-La Roche AG | High temperature reverse transcriptases |
DE4124286A1 (en) * | 1991-07-22 | 1993-01-28 | Boehringer Mannheim Gmbh | METHOD FOR STABILIZING PROTEINS IN OPTICAL TESTS |
FR2690691B1 (en) * | 1992-04-29 | 1999-02-12 | Bio Merieux | METHOD OF AMPLIFYING RNA REQUIRING A SINGLE HANDLING STAGE. |
CA2141430C (en) * | 1992-08-04 | 2009-05-12 | Sherrol H. Mcdonough | Nucleic acid sequence amplification |
WO1994012657A1 (en) * | 1992-11-20 | 1994-06-09 | Katcher Harold L | Direct-phenol pcr, rt and rt-pcr methods |
DE4239969A1 (en) * | 1992-11-27 | 1994-06-01 | Boehringer Mannheim Gmbh | Process for the stabilization of proteins |
EP0655091A1 (en) * | 1993-05-06 | 1995-05-31 | Baxter Diagnostics Inc. | Human papillomavirus detection assay |
WO1995014770A1 (en) * | 1993-11-25 | 1995-06-01 | Pacific Enzymes (1993) Limited | Improved polymerase |
DE69531487T2 (en) * | 1994-01-31 | 2004-06-17 | The Regents Of The University Of California, Oakland | METHOD FOR ELIMINATING SEQUENCING ARTIFACTS |
DE4411588C1 (en) * | 1994-03-30 | 1995-09-28 | Deutsches Rheuma Forschungszen | Buffer, contg. betaine, for RNA- and DNA-polymerase reactions |
US5556771A (en) * | 1995-02-10 | 1996-09-17 | Gen-Probe Incorporated | Stabilized compositions of reverse transcriptase and RNA polymerase for nucleic acid amplification |
-
1997
- 1997-07-23 US US08/899,392 patent/US6013488A/en not_active Expired - Lifetime
- 1997-07-24 DE DE69734913T patent/DE69734913T2/en not_active Expired - Lifetime
- 1997-07-24 EP EP02013871A patent/EP1243661B1/en not_active Expired - Lifetime
- 1997-07-24 DE DE69738354T patent/DE69738354D1/en not_active Expired - Lifetime
- 1997-07-24 EP EP97112672A patent/EP0821059B8/en not_active Expired - Lifetime
- 1997-07-24 CA CA002211367A patent/CA2211367C/en not_active Expired - Fee Related
-
1999
- 1999-10-08 US US09/414,531 patent/US6221599B1/en not_active Expired - Lifetime
-
2001
- 2001-03-13 US US09/803,952 patent/US6372437B2/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
US20010012617A1 (en) | 2001-08-09 |
DE69734913D1 (en) | 2006-01-26 |
US6013488A (en) | 2000-01-11 |
DE69734913T2 (en) | 2006-08-24 |
DE69738354D1 (en) | 2008-01-17 |
EP0821059A2 (en) | 1998-01-28 |
EP0821059B8 (en) | 2006-03-08 |
EP0821059B1 (en) | 2005-12-21 |
EP1243661A1 (en) | 2002-09-25 |
CA2211367C (en) | 2008-04-22 |
US6372437B2 (en) | 2002-04-16 |
US6221599B1 (en) | 2001-04-24 |
EP0821059A3 (en) | 1998-04-22 |
EP1243661B1 (en) | 2007-12-05 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request | ||
MKLA | Lapsed |
Effective date: 20140724 |