WO2012124965A3 - 다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법 - Google Patents

다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법 Download PDF

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Publication number
WO2012124965A3
WO2012124965A3 PCT/KR2012/001808 KR2012001808W WO2012124965A3 WO 2012124965 A3 WO2012124965 A3 WO 2012124965A3 KR 2012001808 W KR2012001808 W KR 2012001808W WO 2012124965 A3 WO2012124965 A3 WO 2012124965A3
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WIPO (PCT)
Prior art keywords
nucleic acid
multiple target
acid sequence
target loci
sequence
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Ceased
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PCT/KR2012/001808
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English (en)
French (fr)
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WO2012124965A2 (ko
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방두희
한효준
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Industry Academic Cooperation Foundation of Yonsei University
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Industry Academic Cooperation Foundation of Yonsei University
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Priority to US14/239,785 priority Critical patent/US20140315202A1/en
Publication of WO2012124965A2 publication Critical patent/WO2012124965A2/ko
Publication of WO2012124965A3 publication Critical patent/WO2012124965A3/ko
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • 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
    • C12Q1/6858—Allele-specific amplification
    • 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
    • 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
    • C12Q1/6869—Methods for sequencing

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

Abstract

본 발명은 다중 타겟 위치(multiple target loci)를 하나의 어셈블된 핵산서열로의 어셈블리하는 방법 및 이를 이용한 동시 검출방법에 관한 것이다. 본 발명의 방법은 두 번의 PCR, 즉 제1차 PCR(polymerase chain reaction) 및 제2차 PCR 반응을 통해 다중 타겟 위치를 하나의 어셈블된 핵산서열로 어셈블리시킨다. 보다 상세하게는, 본 발명에서 이용되는 제1차 증폭용 프라이머쌍(정방향 프라이머 및 역방향 프라이머)은 타겟-특이적 서열(타겟 혼성화 뉴클레오타이드 서열) 및 5'-플랭킹 어셈블리 스페이서 서열(오버래핑 서열)로 이루어져 있다. 또한, 상기 제1차 증폭용 프라이머쌍을 이용하여 증폭된 제1차 증폭결과물을 제2차 증폭용 프라이머 세트를 통해 간편하고 용이하게 하나의 단축된 핵산서열로 어셈블리시켜 다중 타겟 위치를 동시에 검출할 수 있다. 따라서, 본 발명의 방법 및 키트는 시료(바람직하게는, 인간의 혈액)의 DNA 서열 상의 다중 변이성(예컨대, SNPs)을 동시에 검출 및 분석함으로써, 변이 검출을 위한 시퀀싱 비용을 현저하게 감소시킬 수 있을 뿐 아니라 맞춤형 의약의 개념을 실현하기 위한 중요한 접근방법 및 수단을 제공한다.
PCT/KR2012/001808 2011-03-16 2012-03-13 다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법 Ceased WO2012124965A2 (ko)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US14/239,785 US20140315202A1 (en) 2011-03-16 2012-03-13 Method for assembling multiple target loci to single nucleic acid sequence

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR10-2011-0023184 2011-03-16
KR1020110023184A KR101306988B1 (ko) 2011-03-16 2011-03-16 다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법

Publications (2)

Publication Number Publication Date
WO2012124965A2 WO2012124965A2 (ko) 2012-09-20
WO2012124965A3 true WO2012124965A3 (ko) 2012-12-27

Family

ID=46831198

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2012/001808 Ceased WO2012124965A2 (ko) 2011-03-16 2012-03-13 다중 타겟 위치의 단일 핵산서열로의 어셈블리 방법

Country Status (3)

Country Link
US (1) US20140315202A1 (ko)
KR (1) KR101306988B1 (ko)
WO (1) WO2012124965A2 (ko)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6204025B1 (en) * 1997-09-29 2001-03-20 City Of Hope Efficient linking of nucleic acid segments
US20090042258A1 (en) * 2002-04-12 2009-02-12 New England Biolabs, Inc. Methods and Compositions for DNA Manipulation
US7498131B2 (en) * 2001-12-14 2009-03-03 Keygene, Nv Analysis and detection of multiple target sequences using circular probes
US20100143900A1 (en) * 2008-12-04 2010-06-10 Peluso Paul S Asynchronous sequencing of biological polymers

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6124092A (en) * 1996-10-04 2000-09-26 The Perkin-Elmer Corporation Multiplex polynucleotide capture methods and compositions

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6204025B1 (en) * 1997-09-29 2001-03-20 City Of Hope Efficient linking of nucleic acid segments
US7498131B2 (en) * 2001-12-14 2009-03-03 Keygene, Nv Analysis and detection of multiple target sequences using circular probes
US20090042258A1 (en) * 2002-04-12 2009-02-12 New England Biolabs, Inc. Methods and Compositions for DNA Manipulation
US20100143900A1 (en) * 2008-12-04 2010-06-10 Peluso Paul S Asynchronous sequencing of biological polymers

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HAN, H. ET AL.: "Multiple target loci assembly sequencing (mTAS)", ANALYTICAL BIOCHEMISTRY, vol. 415, 16 April 2011 (2011-04-16), pages 218 - 220 *

Also Published As

Publication number Publication date
KR101306988B1 (ko) 2013-09-10
WO2012124965A2 (ko) 2012-09-20
US20140315202A1 (en) 2014-10-23
KR20120105640A (ko) 2012-09-26

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