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1. WO1996041005 - METHODS FOR NUCLEIC ACID DETECTION, SEQUENCING, AND CLONING USING EXONUCLEASE

Publication Number WO/1996/041005
Publication Date 19.12.1996
International Application No. PCT/US1996/009208
International Filing Date 06.06.1996
IPC
C12N 9/22 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes, e.g. ligases (6.); Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating, or purifying enzymes
14Hydrolases (3.)
16acting on ester bonds (3.1)
22Ribonucleases
C12Q 1/68 2006.01
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING 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
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
68involving nucleic acids
CPC
C07K 14/005
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
005from viruses
C07K 14/35
CCHEMISTRY; METALLURGY
07ORGANIC CHEMISTRY
KPEPTIDES
14Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
195from bacteria
35from Mycobacteriaceae (F)
C12N 2750/14222
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
2750ssDNA Viruses
00011ssDNA Viruses
14011Parvoviridae
14211Erythrovirus, e.g. B19 virus
14222New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
C12N 9/22
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
9Enzymes; Proenzymes; Compositions thereof
14Hydrolases (3)
16acting on ester bonds (3.1)
22Ribonucleases ; RNAses, DNAses
C12Q 1/6806
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6806Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 1/6827
CCHEMISTRY; METALLURGY
12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS
1Measuring or testing processes involving enzymes, nucleic acids or microorganisms
68involving nucleic acids
6813Hybridisation assays
6827for detection of mutation or polymorphism
Applicants
  • MURTAGH, James, J. [US]/[US]
  • THUNNISSEN, Frederik, B., J., M. [NL]/[NL]
Inventors
  • MURTAGH, James, J.
  • THUNNISSEN, Frederik, B., J., M.
Agents
  • PERRYMAN, David, G.
Priority Data
08/479,72307.06.1995US
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) METHODS FOR NUCLEIC ACID DETECTION, SEQUENCING, AND CLONING USING EXONUCLEASE
(FR) PROCEDES DE DETECTION, DE SEQUENÇAGE ET DE CLONAGE D'ACIDES NUCLEIQUES A L'AIDE DE L'EXONUCLEASE
Abstract
(EN)
The present invention provides a method of detecting the presence of a nucleotide sequence within a double-stranded DNA in a sample comprising: (a) digesting the double-stranded DNA with an exonuclease which converts at least a portion of the double-stranded DNA to single-stranded DNA; (b) hybridizing the single-stranded DNA with i) a first nucleic acid probe adapted with a moiety which can be captured to a solid support, and ii) a second nucleic acid probe labeled with a detectable moiety which can hybridize with the single-stranded DNA adjacent the hybridized first nucleic acid probe; (c) ligating the hybridized first and second nucleic acid probes; (d) capturing the moiety on the first nucleic acid probe hybridized to the DNA to the solid support; (e) denaturing the ligated first and second nucleic acid probes from the hybridized single-stranded DNA; (f) removing uncaptured labeled probe; and, (g) detecting the presence of captured detectable moiety, the presence of captured detectable moiety indicating the presence of the nucleotide sequence within the double-stranded DNA in the sample.
(FR)
Procédé de détection de la présence d'une séquence nucléotidique dans un ADN à double brin d'un échantillon qui consiste (a) à faire digérer l'ADN à double brin par une exonucléase qui convertit au moins une partie de l'ADN à double brin en ADN à simple brin, (b) à hybrider l'ADN à simple brin avec i) une première sonde d'acide nucléique adaptée à l'aide d'une fraction qui peut être fixée à un support solide et ii) une deuxième sonde d'acide nucléique marquée à l'aide d'une fraction détectable qui peut s'hybrider avec l'ADN à simple brin adjacent à la première sonde d'acide nucléique hybridée, (c) à ligaturer les première et deuxième sondes d'acide nucléique hybridées, (d) à fixer la fraction sur la première sonde d'acide nucléique hybridée à l'ADN au support solide, (e) à dénaturer les première et deuxième sondes d'acide nucléique ligaturées de l'ADN à simple brin hybridé, (f) à éliminer la sonde marquée non fixée et (g) à détecter la présence de la fraction détectable fixée indiquant la présence de la séquence nucléotidique au sein de l'ADN à double brin de l'échantillon.
Also published as
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