WO2001029248B1 - Method for amplifying and detecting nucleic acid - Google Patents

Method for amplifying and detecting nucleic acid

Info

Publication number
WO2001029248B1
WO2001029248B1 PCT/KR1999/000628 KR9900628W WO0129248B1 WO 2001029248 B1 WO2001029248 B1 WO 2001029248B1 KR 9900628 W KR9900628 W KR 9900628W WO 0129248 B1 WO0129248 B1 WO 0129248B1
Authority
WO
WIPO (PCT)
Prior art keywords
method according
dttp
dctp
set
datp
Prior art date
Application number
PCT/KR1999/000628
Other languages
French (fr)
Other versions
WO2001029248A3 (en
WO2001029248A2 (en
Inventor
Gi Young Jang
Original Assignee
Bionex Inc
Gi Young Jang
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 Bionex Inc, Gi Young Jang filed Critical Bionex Inc
Priority to PCT/KR1999/000628 priority Critical patent/WO2001029248A2/en
Publication of WO2001029248A2 publication Critical patent/WO2001029248A2/en
Publication of WO2001029248A3 publication Critical patent/WO2001029248A3/en
Publication of WO2001029248B1 publication Critical patent/WO2001029248B1/en

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844Nucleic acid amplification reactions
    • C12Q1/686Polymerase chain reaction [PCR]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869Methods for sequencing
    • C12Q1/6874Methods for sequencing involving nucleic acid arrays, e.g. sequencing by hybridisation

Abstract

The present invention provides a method for amplifying and detecting target nucleic acid. One method of present invention comprises Polymerase Chain Reaction (PCR) amplification process with two different standard oligonucleotide primers along with extra number of oligonucleotide primers bound to a solid support. The extra number of oligonucleotide primers bound to the solid support can be co-amplified along with equal or preferably unequal amount of two standard oligonucleotide primers. Another method of present invention involves multi-number of target amplification by periodical addition of the oligonucleotide primers during the amplification process. These processes are useful for specifying the amplified nucleic acid, which allow the simple and convenient format for diagnostic and DNA sequencing.

Claims

AMENDED CLAIMS
[received by the International Bureau on 16 April 2001 (16 04 01), original claims 21-23 amended, remaining claims unchanged (1 page)] acids,
(0 repeating the steps (c) through (e) at least once, and
(g) repeating the steps (b) through (f) at least once
1 5 method according to claim 14, wherein the size of the ohgonucleotide -> pnmer is fi om 7 to 100
16 A method according to claim 14, wherein the steps (b) and (c) are perfoπned at the temperature of 85-98 °C
17 A method according to claim 14, wherein the step (d) is performed at the tempeiature of 10-75 °C 0 I S A method according to claim 14, w herein the step (e) is performed at the lempciJtui e of 20-75 °C
19 A method according to claim 14, wherein in the step (b) the mixture of four diffei ent nucleoside triphosphates and enzyme are further added
20 A method according to claim 14, wherein in the step (a), the reaction mixture 5 further contains plural oligonucleotide pnmers bound to at least one solid support
21. A method according to claim 14, wherein in the step (b), the oligonucleotide pi uneis bound to the solid support are further added
22 A method according to claim 14. wherein the two standard ohgonucleotide pnmers are selected respectively from the group consisting of a unlabeled 0 oix'onuclcotide pπmer, a 5'end fluoiescenth labeleα ohgonucleotide pπmer, a 5'end Biotin labeled ohgonucleotide primer, α 5'end radioisotope-labeled o gonucleotide pπinei and a 5'end Digoxigemn-labeled oligonucletide primer
23 A method according to claim 14, w Herein the four different nucleoside tnphosphates are selected from the group consisting of the set of dATP, dTTP, dGTP and dCTP, the set of dTTP, dGTP, dCTP and fluorescentlv-labeled dATP, the set of d VTP, dTTP, dCTP, and 7-Deaza-dGTP, the set of dTTP. dCTP, 7-Deaza-dGTP and fluorescetly-labeled dATP, the set of dATP, dTTP. dCTP, and digoxtgenin-dUTP, the set of dATP, dTTP, dCTP and fluorescently label dUTP and the set of dATP, dTTP, dCTP and biotin label dUTP 0 24 A method according to claim 14, wherein the steps (a) through
Figure imgf000002_0001
are performed by using a robotic workstation having an automatic multi-pipetting function
18
PCT/KR1999/000628 1999-10-19 1999-10-19 Method for amplifying and detecting nucleic acid WO2001029248A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/KR1999/000628 WO2001029248A2 (en) 1999-10-19 1999-10-19 Method for amplifying and detecting nucleic acid

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/KR1999/000628 WO2001029248A2 (en) 1999-10-19 1999-10-19 Method for amplifying and detecting nucleic acid

Publications (3)

Publication Number Publication Date
WO2001029248A2 WO2001029248A2 (en) 2001-04-26
WO2001029248A3 WO2001029248A3 (en) 2001-09-20
WO2001029248B1 true WO2001029248B1 (en) 2001-12-06

Family

ID=19571119

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR1999/000628 WO2001029248A2 (en) 1999-10-19 1999-10-19 Method for amplifying and detecting nucleic acid

Country Status (1)

Country Link
WO (1) WO2001029248A2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102005029810B4 (en) * 2005-06-27 2008-11-13 Siemens Ag A method for detecting nucleotide sequences, using the method and test kit

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE69029726D1 (en) * 1989-03-22 1997-02-27 Cemu Bioteknik Ab Solid phase diagnosis of medical conditions
WO1995006753A1 (en) * 1993-09-03 1995-03-09 The United States of America, represented by The Secretary, Department of Health & Human Services Method and compositions for primer specific solid-phase detection of pcr products
WO1995015400A1 (en) * 1993-12-03 1995-06-08 The Johns Hopkins University Genotyping by simultaneous analysis of multiple microsatellite loci
FR2726286B1 (en) * 1994-10-28 1997-01-17 Genset Sa Method for nucleic acid amplification in the solid phase and reagents useful kit for carrying out this method
US5693467A (en) * 1995-05-19 1997-12-02 The American Type Culture Collection Mycoplasma polymerase chain reaction testing system using a set of mixed and single sequence primers
DE19811731A1 (en) * 1998-03-18 1999-09-23 November Ag Molekulare Medizin Detecting nucleic acid by amplification, with use of third, immobilized primer

Also Published As

Publication number Publication date
WO2001029248A3 (en) 2001-09-20
WO2001029248A2 (en) 2001-04-26

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