WO2002050094A3 - Nucleic acid synthesis by primer extension on successive templates - Google Patents

Nucleic acid synthesis by primer extension on successive templates Download PDF

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Publication number
WO2002050094A3
WO2002050094A3 PCT/GB2001/005703 GB0105703W WO0250094A3 WO 2002050094 A3 WO2002050094 A3 WO 2002050094A3 GB 0105703 W GB0105703 W GB 0105703W WO 0250094 A3 WO0250094 A3 WO 0250094A3
Authority
WO
WIPO (PCT)
Prior art keywords
strand
template
dna sequence
template oligonucleotides
nucleic acid
Prior art date
Application number
PCT/GB2001/005703
Other languages
French (fr)
Other versions
WO2002050094A2 (en
Inventor
Mark Mcalister
Renos Savva
Uma Bhattacharyya
Original Assignee
Birkbeck College
Mark Mcalister
Renos Savva
Uma Bhattacharyya
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
Application filed by Birkbeck College, Mark Mcalister, Renos Savva, Uma Bhattacharyya filed Critical Birkbeck College
Priority to AU2002216229A priority Critical patent/AU2002216229A1/en
Publication of WO2002050094A2 publication Critical patent/WO2002050094A2/en
Publication of WO2002050094A3 publication Critical patent/WO2002050094A3/en

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    • 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
    • C12N15/102—Mutagenizing nucleic acids
    • C12N15/1031—Mutagenizing nucleic acids mutagenesis by gene assembly, e.g. assembly by oligonucleotide extension PCR
    • 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
    • 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

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Plant Pathology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Chemical Kinetics & Catalysis (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 relates to methods and kits for synthesizing a DNA sequence which employ a series of overlapping template oligonucleotides having sequences corresponding to one strand of a DNA sequence, these template oligonucleotides being incapable of extension in the synthesis reaction. The method uses one primer that primes synthesis of one strand of the DNA sequence, using a series of template oligonucleotides corresponding solely to the complementary strand as template. A second primer that primes synthesis of the complementary strand using the newly synthesised strand as template can be used. In one embodiment, the template oligonucleotides have modified 3' termini to prevent DNA polymerase mediated strand extension and nucleotide backbone modifications between one or more 3' nucleotides, to inhibit or prevent removal of the 3' modified nucleotides by the 3'-5' exonuclease activity of proof-reading polymerases, i.e. to confer some resistance to the exonuclease.
PCT/GB2001/005703 2000-12-20 2001-12-20 Nucleic acid synthesis by primer extension on successive templates WO2002050094A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU2002216229A AU2002216229A1 (en) 2000-12-20 2001-12-20 Nucleic acid synthesis by primer extension on successive templates

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0031093.8 2000-12-20
GB0031093A GB0031093D0 (en) 2000-12-20 2000-12-20 Materials and methods for nucleic acid systhesis

Publications (2)

Publication Number Publication Date
WO2002050094A2 WO2002050094A2 (en) 2002-06-27
WO2002050094A3 true WO2002050094A3 (en) 2003-04-24

Family

ID=9905486

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2001/005703 WO2002050094A2 (en) 2000-12-20 2001-12-20 Nucleic acid synthesis by primer extension on successive templates

Country Status (3)

Country Link
AU (1) AU2002216229A1 (en)
GB (1) GB0031093D0 (en)
WO (1) WO2002050094A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005017153A2 (en) * 2003-08-04 2005-02-24 Blue Heron Biotechnology, Inc. Methods for synthesis of defined polynucleotides

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5302509A (en) * 1989-08-14 1994-04-12 Beckman Instruments, Inc. Method for sequencing polynucleotides
WO1999014370A1 (en) * 1997-09-15 1999-03-25 Hiatt Andrew C De novo polynucleotide synthesis using rolling templates
WO2000029616A1 (en) * 1998-11-12 2000-05-25 The Perkin-Elmer Corporation Ligation assembly and detection of polynucleotides on solid-support

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5302509A (en) * 1989-08-14 1994-04-12 Beckman Instruments, Inc. Method for sequencing polynucleotides
WO1999014370A1 (en) * 1997-09-15 1999-03-25 Hiatt Andrew C De novo polynucleotide synthesis using rolling templates
WO2000029616A1 (en) * 1998-11-12 2000-05-25 The Perkin-Elmer Corporation Ligation assembly and detection of polynucleotides on solid-support

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PRODROMOU CHRISOSTOMOS ET AL: "Recursive PCR: A novel technique for total gene synthesis", PROTEIN ENGINEERING, OXFORD UNIVERSITY PRESS, SURREY, GB, vol. 5, no. 8, 1992, pages 827 - 829, XP002176276, ISSN: 0269-2139 *

Also Published As

Publication number Publication date
WO2002050094A2 (en) 2002-06-27
AU2002216229A1 (en) 2002-07-01
GB0031093D0 (en) 2001-01-31

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