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WO2012115464A3 - Composition for hot-start pcr comprising blocking oligonucleotide - Google Patents

Composition for hot-start pcr comprising blocking oligonucleotide Download PDF

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Publication number
WO2012115464A3
WO2012115464A3 PCT/KR2012/001387 KR2012001387W WO2012115464A3 WO 2012115464 A3 WO2012115464 A3 WO 2012115464A3 KR 2012001387 W KR2012001387 W KR 2012001387W WO 2012115464 A3 WO2012115464 A3 WO 2012115464A3
Authority
WO
WIPO (PCT)
Prior art keywords
composition
hot
pcr
primer
blocking oligonucleotide
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.)
Ceased
Application number
PCT/KR2012/001387
Other languages
French (fr)
Other versions
WO2012115464A2 (en
Inventor
Jun Hee Lee
So Ra Choi
Nam Il Kim
Han Oh Park
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Bioneer Corp
Original Assignee
Bioneer Corp
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 Bioneer Corp filed Critical Bioneer Corp
Publication of WO2012115464A2 publication Critical patent/WO2012115464A2/en
Publication of WO2012115464A3 publication Critical patent/WO2012115464A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12Q1/686—Polymerase chain reaction [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
    • 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
    • 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
    • C12Q2527/00—Reactions demanding special reaction conditions
    • C12Q2527/101—Temperature
    • 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
    • C12Q2549/00—Reactions characterised by the features used to influence the efficiency or specificity
    • C12Q2549/10—Reactions characterised by the features used to influence the efficiency or specificity the purpose being that of reducing false positive or false negative signals
    • C12Q2549/101—Hot start

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)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (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)

Abstract

The present invention relates to a composition for hot-start PCR, more precisely a composition for hot-start PCR characteristically prepared by adding the blocking oligonucleotide having the hydroxyl group at 3'-end blocked and the nucleotide sequence complementary to the primer to the conventional PCR composition containing reaction buffer, MgCl2, 4 kinds of dNTP, and DNA polymerase. The composition for hot-start PCR of the present invention can give more authentic PCR results by inhibiting smear band of PCR amplification product caused by non-specific reaction or primer-dimer formation at low temperature since the blocking oligonucleotide included therein can be dissociated at the lower temperature than melting temperature of the primer.
PCT/KR2012/001387 2011-02-25 2012-02-23 Composition for hot-start pcr comprising blocking oligonucleotide Ceased WO2012115464A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2011-0017226 2011-02-25
KR20110017226A KR101424192B1 (en) 2011-02-25 2011-02-25 Composition for Hot-start PCR Comprising Blocking Oligonucleotide

Publications (2)

Publication Number Publication Date
WO2012115464A2 WO2012115464A2 (en) 2012-08-30
WO2012115464A3 true WO2012115464A3 (en) 2012-12-20

Family

ID=46721349

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2012/001387 Ceased WO2012115464A2 (en) 2011-02-25 2012-02-23 Composition for hot-start pcr comprising blocking oligonucleotide

Country Status (2)

Country Link
KR (1) KR101424192B1 (en)
WO (1) WO2012115464A2 (en)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102124058B1 (en) 2013-09-16 2020-06-17 삼성전자주식회사 Polynucleotide and use thereof
WO2015063154A1 (en) * 2013-10-31 2015-05-07 Multiplicom N.V. Blocking primers in multiplex pcr based assays
LT3402880T (en) 2016-01-15 2024-03-12 Thermo Fisher Scientific Baltics Uab Thermophilic dna polymerase mutants
EP3402826A1 (en) 2016-01-15 2018-11-21 Thermo Fisher Scientific Baltics UAB Antibodies that bind thermophilic dna polymerases
KR101897490B1 (en) 2016-12-02 2018-09-12 건국대학교 산학협력단 A composition for PCR containing a polyethyleneglycol-engrafted nano-sized graphene oxide
US11618891B2 (en) 2017-06-26 2023-04-04 Thermo Fisher Scientific Baltics Uab Thermophilic DNA polymerase mutants
KR102141312B1 (en) 2019-04-19 2020-08-04 주식회사 제노헬릭스 Small RNA-primed Xenosensor module amplification mediated small RNA detection method
GB202012480D0 (en) * 2020-08-11 2020-09-23 Univ Oxford Innovation Ltd Switch Oligonucleotide
DE202020004407U1 (en) * 2020-10-20 2020-11-24 Friz Biochem Gmbh Functionalized microtiter plate, reaction solution, and kit for preparing a reaction solution
KR20250000081A (en) 2023-06-23 2025-01-02 고려대학교 산학협력단 Development of a Simple Hot-start PCR using Escherichia coli expressing Taq DNA polymerase

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ASHRAFI, E. H. ET AL.: "Improved PCR specificity with Hot Start PCR primers.", BIOTECHNIQUES., vol. 47, no. 3, September 2009 (2009-09-01), pages 789 - 790 *
CHOU, Q. ET AL.: "Prevention of pre-PCR mis-priming and primer dimerization improves low-copy-number amplifications.", NUCLEIC ACIDS RES., vol. 20, no. 7, 11 April 1992 (1992-04-11), pages 1717 - 1723 *
CRADIC, K. W. ET AL.: "Substitution of 3'-phosphate cap with a carbon-based blocker reduces the possibility of fluorescence resonance energy transfer probe failure in real-time PCR assays.", CLINICAL CHEMISTRY, vol. 50, no. 6, June 2004 (2004-06-01), pages 1080 - 1082 *
LEBEDEV, A. V. ET AL.: "Hot start PCR with heat-activatable primers: a novel approach for improved PCR performance.", NUCLEIC ACIDS RESEARCH, vol. 36, no. 20, November 2008 (2008-11-01), pages E131 *
VESTHEIM, H. ET AL.: "Blocking primers to enhance PCR amplification of rare sequences in mixed samples- a case study on prey DNA in Antarctic krill stomachs.", FRONTIERS IN ZOOLOGY., vol. 5, no. 12, 20 July 2008 (2008-07-20) *

Also Published As

Publication number Publication date
KR20120097793A (en) 2012-09-05
KR101424192B1 (en) 2014-07-28
WO2012115464A2 (en) 2012-08-30

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