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WO2003012066A3 - Magnesium precipitate hot start method for molecular manipulation of nucleic acids - Google Patents

Magnesium precipitate hot start method for molecular manipulation of nucleic acids Download PDF

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Publication number
WO2003012066A3
WO2003012066A3 PCT/US2002/024533 US0224533W WO03012066A3 WO 2003012066 A3 WO2003012066 A3 WO 2003012066A3 US 0224533 W US0224533 W US 0224533W WO 03012066 A3 WO03012066 A3 WO 03012066A3
Authority
WO
WIPO (PCT)
Prior art keywords
magnesium
nucleic acids
hot start
start method
kits
Prior art date
Application number
PCT/US2002/024533
Other languages
French (fr)
Other versions
WO2003012066A2 (en
Inventor
Wayne M Barnes
Katherine R Rowlyk
Original Assignee
Wayne M Barnes
Katherine R Rowlyk
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.)
Filing date
Publication date
Priority claimed from US09/920,872 external-priority patent/US6403341B1/en
Application filed by Wayne M Barnes, Katherine R Rowlyk filed Critical Wayne M Barnes
Priority to EP02763410A priority Critical patent/EP1419275A4/en
Priority to AU2002327421A priority patent/AU2002327421A1/en
Publication of WO2003012066A2 publication Critical patent/WO2003012066A2/en
Publication of WO2003012066A3 publication Critical patent/WO2003012066A3/en

Links

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
    • 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
    • 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
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • 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
    • C12Q1/686—Polymerase chain reaction [PCR]

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Analytical Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The present invention provides methods of performing enzymatic reactions, including PCR, which require the use of magnesium dependent enzymes, including restriction endonucleases, ligases, and reverse transcriptases. The method is based on sequestration of magnesium ions in the form of a precipitate which renders a magnesium dependent enzyme inactive until the appropriate time in the reaction when a certain temperature is reached and the magnesium ions are released from the precipitate. Also provided are kits comprising reagents and instructions for amplifying a target nucleic acid, for DNA digestion and ligation, and for reverse transcription of RNA into cDNA. Furthermore, the kits and reagents of the present invention can be utilized in other reactions requiring magnesium dependent enzymes.
PCT/US2002/024533 2001-08-02 2002-08-02 Magnesium precipitate hot start method for molecular manipulation of nucleic acids WO2003012066A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP02763410A EP1419275A4 (en) 2001-08-02 2002-08-02 Magnesium precipitate hot start method for molecular manipulation of nucleic acids
AU2002327421A AU2002327421A1 (en) 2001-08-02 2002-08-02 Magnesium precipitate hot start method for molecular manipulation of nucleic acids

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US30964601P 2001-08-02 2001-08-02
US09/920,872 2001-08-02
US60/309,646 2001-08-02
US09/920,872 US6403341B1 (en) 2001-08-02 2001-08-02 Magnesium precipitate hot start method for PCR
US10/091,784 US20030082567A1 (en) 2001-08-02 2002-03-06 Mineral hot start PCR method
US10/091,784 2002-03-06

Publications (2)

Publication Number Publication Date
WO2003012066A2 WO2003012066A2 (en) 2003-02-13
WO2003012066A3 true WO2003012066A3 (en) 2003-12-04

Family

ID=27376974

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2002/024533 WO2003012066A2 (en) 2001-08-02 2002-08-02 Magnesium precipitate hot start method for molecular manipulation of nucleic acids

Country Status (3)

Country Link
EP (1) EP1419275A4 (en)
AU (1) AU2002327421A1 (en)
WO (1) WO2003012066A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9562263B2 (en) 2007-07-14 2017-02-07 Ionian Technologies, Inc. Nicking and extension amplification reaction for the exponential amplification of nucleic acids

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10315640A1 (en) 2003-04-04 2004-10-14 Ignatov, Konstantin Process for the controlled release of components into a solution
EP3778915A1 (en) * 2012-06-08 2021-02-17 Ionian Technologies, LLC Nucleic acid amplifications
WO2015113828A1 (en) 2014-01-31 2015-08-06 Qiagen Gmbh Cation chelator hot start
WO2019118652A1 (en) 2017-12-12 2019-06-20 Essenlix Corporation Sample manipulation and assay with rapid temperature change

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE33487E (en) * 1986-03-12 1990-12-11 Palintest Limited Water treatment and soil testing
WO1996000301A1 (en) * 1994-06-24 1996-01-04 The University Of Houston Encapsulated pcr reagents
US5565339A (en) * 1992-10-08 1996-10-15 Hoffmann-La Roche Inc. Compositions and methods for inhibiting dimerization of primers during storage of polymerase chain reaction reagents
US5599660A (en) * 1993-01-19 1997-02-04 Pharmacia Biotech Inc. Method and preparation for sequential delivery of wax-embedded, inactivated biological and chemical reagents
US6214557B1 (en) * 2000-06-06 2001-04-10 Washington University Cold sensitive mutant DNA polymerases

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5250429A (en) * 1991-09-20 1993-10-05 Pharmacia P-L Biochemicals Inc. Glassified restriction enzymes

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
USRE33487E (en) * 1986-03-12 1990-12-11 Palintest Limited Water treatment and soil testing
US5565339A (en) * 1992-10-08 1996-10-15 Hoffmann-La Roche Inc. Compositions and methods for inhibiting dimerization of primers during storage of polymerase chain reaction reagents
US5599660A (en) * 1993-01-19 1997-02-04 Pharmacia Biotech Inc. Method and preparation for sequential delivery of wax-embedded, inactivated biological and chemical reagents
WO1996000301A1 (en) * 1994-06-24 1996-01-04 The University Of Houston Encapsulated pcr reagents
US6214557B1 (en) * 2000-06-06 2001-04-10 Washington University Cold sensitive mutant DNA polymerases

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1419275A4 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9562263B2 (en) 2007-07-14 2017-02-07 Ionian Technologies, Inc. Nicking and extension amplification reaction for the exponential amplification of nucleic acids
US9562264B2 (en) 2007-07-14 2017-02-07 Ionian Technologies, Inc. Nicking and extension amplification reaction for the exponential amplification of nucleic acids

Also Published As

Publication number Publication date
WO2003012066A2 (en) 2003-02-13
EP1419275A2 (en) 2004-05-19
AU2002327421A1 (en) 2003-02-17
EP1419275A4 (en) 2007-05-30

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