A method for selectively detecting subsets of nucleic acid...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2562288

Method for selectively detecting nucleic acid molecules comprising structural aberrations that are capable of being converted into nicks comprising generating linear nucleic acids from a selected nucleic acid substrate population; denaturing and re-annealing the linear nucleic acids to form nucleic acid duplexes; masking the nucleic acid duplex termini and internal structural aberrations with a masking component; modifying the masked nucleic acids by introducing nicks therein using at least an enzyme possessing endonuclease activity;labelling the modified nucleic acids with labelled nucleotides via nucleic acid nick translation with at least an enzyme displaying a nucleic acid polymerase activity; and selecting and identifying the labelled nucleic acid.

L'invention concerne un procédé permettant la détection sélective de molécules d'acide nucléique contenant des aberrations de structure pouvant être converties en coupures, ce procédé consistant à générer des acides nucléiques linéaires à partir d'une population choisie de substrat nucléotidique, à dénaturer et à renaturer les acides nucléiques linéaires afin de former des acides nucléiques double brin, à masquer les terminus des acides nucléiques double brin et les aberrations de structure internes au moyen d'un composant de masquage, à modifier les acides nucléiques masqués par l'introduction de coupures dans ces derniers, en utilisant au moins une enzyme possédant une activité endonucléase, à marquer les acides nucléiques modifiés avec des nucléotides marqués, par l'intermédiaire d'une translation de coupure d'acide nucléique, au moyen d'au moins une enzyme présentant une activité polymérase d'acide nucléique, et à sélectionner et à identifier l'acide nucléique ainsi marqué.

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