Method for analysing the length of a nucleic acid molecule

Details Method for analysing the length of a nucleic acid molecule Method for analysing the length of a nucleic acid molecule

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2400339

A method of analysing the length of a particular region within a nucleic acid, said method comprising a) subjecting a sample of said nucleic acid to a plurality of amplification reactions in which said region is amplified, wherein the time of the extension phase in each of said reactions is varied; b) monitoring the progress of said amplification reactions; c) determining the minimum time during which extension phase of the amplification is completed within each reaction mixture and relating that to the length of the sequence undergoing extension. The method, combined with melting point analysis will allow percentage GC content of a sequence to be determined. Length analysis of this type can be used in diagnosis or analysis as well as in recombinant DNA technology to check for the presence of concatamers, and in taxonomic classification or forensics. Apparatus for use in the method is also described and claimed.

L'invention porte sur une méthode d'analyse de la longueur de régions particulières d'un acide nucléique consistant: (a) à soumettre un échantillon dudit acide à une série d'amplifications qui amplifient ladite région, le temps de la phase d'extension de chacune des réactions étant différent; (b) à suivre la progression desdites réactions d'amplification; (c) à déterminer le temps minimum pendant lequel la phase d'extension de l'amplification s'accomplit pour chacun des mélanges réactifs puis à le rapporter à la longueur de la séquence subissant l'extension. Cette méthode, combinée à l'analyse du point de fusion permet de déterminer le pourcentage de GC contenu dans la séquence. Une telle analyse de longueur peut servir pour des diagnostics ou des analyses, et dans le cadre de techniques de recombinaison d'ADN, à vérifier la présence de concatamères, ou pour la classification taxonomique ou en médecine légale. L'invention porte en outre sur un appareil servant à mettre en oeuvre la méthode.

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