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1. WO2009010776 - DETECTION OF METHYLATION IN NUCLEIC ACID SEQUENCE

Publication Number WO/2009/010776
Publication Date 22.01.2009
International Application No. PCT/GB2008/002487
International Filing Date 18.07.2008
IPC
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
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
C12Q 1/686
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
6844Nucleic acid amplification reactions
686Polymerase chain reaction [PCR]
Y10T 436/143333
YSECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
10TECHNICAL SUBJECTS COVERED BY FORMER USPC
TTECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
436Chemistry: analytical and immunological testing
14Heterocyclic carbon compound [i.e. , O, S, N, Se, Te, as only ring hetero atom]
142222Hetero-O [e.g., ascorbic acid, etc.]
143333Saccharide [e.g., DNA, etc.]
Applicants
  • ABERYSTWYTH UNIVERSITY [GB]/[GB] (AllExceptUS)
  • WILKINSON, Michael, James [GB]/[GB] (UsOnly)
  • LOPEZ, Carlos Marcelino Rodriguez [ES]/[GB] (UsOnly)
Inventors
  • WILKINSON, Michael, James
  • LOPEZ, Carlos Marcelino Rodriguez
Agents
  • PILKINGTON, Stephanie
Priority Data
0714058.519.07.2007GB
Publication Language English (EN)
Filing Language English (EN)
Designated States
Title
(EN) DETECTION OF METHYLATION IN NUCLEIC ACID SEQUENCE
(FR) DÉTECTION D'UNE MÉTHYLATION DANS UNE SÉQUENCE D'ACIDE NUCLÉIQUE
Abstract
(EN)
The present invention provides a method for detecting and/or quantifying the presence of, and relative abundance of, methylated nucleic acid bases within double- stranded nucleic acid by i) contacting a double-stranded nucleic acid sample with an intercalating fluorescent dye when bound to the nucleic acid sample fluoresces when exposed to light of a wavelength capable of causing the dye to fluoresce; 2) altering the hybridisation conditions of the solution containing the double-stranded nucleic acid-dye complex such that dissociation of the two strands of the said nucleic acid-dye complex occurs at a rate that permits progressive release of the dye 3) and monitoring the difference in fluorescence, and uses thereof.
(FR)
La présente invention porte sur un procédé pour détecter et/ou quantifier la présence et l'abondance relative de bases d'acide nucléique méthylées dans un acide nucléique double brin par les opérations consistant à i) mettre en contact un échantillon d'acide nucléique double brin avec un colorant fluorescent d'insertion qui, lorsqu'il est lié à l'échantillon d'acide nucléique, émet une fluorescence lorsqu'il est exposé à une lumière d'une longueur d'onde capable d'amener le colorant à émettre une fluorescence; 2) modifier les conditions d'hybridation de la solution contenant le complexe acide nucléique double brin-colorant, de telle sorte qu'une dissociation des deux brins de ce complexe acide nucléique colorant se produit à une vitesse qui permet une libération progressive du colorant; et 3) surveiller la différence de fluorescence. L'invention porte également sur des utilisations de ce procédé.
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