Methods and compositions for genotyping

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 21/00 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2497570

The invention provides compositions and methods for performing primer extension reactions, including employment of amplification primers having 5' tags to incorporate into amplicons variant nucleotides of interest from target nucleic acids at known ratios, with or without sequences surrounding the variant nucleotides of interest. The invention provides identifying the variant nucleotides generated from the target nucleic acid and generated from the 5' tags, comparing the results, evaluating the efficiency of the primer extension reactions, and monitoring the efficacy of such reactions. The invention accounts for DNA sequence and experimental variables that may affect efficiency of incorporation of nucleotides, and provides a reference point for the interpretation of polymorphisms. The invention also provides methods of breeding scrapie-resistant sheep populations.

L'invention concerne des compositions et des méthodes permettant d'exécuter des réactions d'extension d'amorce, dont l'utilisation d'amorces d'amplification avec étiquettes 5' pour incorporation dans des amplicons de nucléotides variants d'intérêt à partir d'acides nucléiques cibles à des taux connus, avec ou sans séquences entourant les nucléotides variants d'intérêt. Cette invention permet d'identifier les nucléotides variants obtenus à partir de l'acide nucléique cible et des étiquettes 5', de comparer les résultats, d'évaluer l'efficacité des réactions d'extension d'amorce et de surveiller l'efficience de telles réactions. L'invention prend en compte une séquence d'ADN et des variables expérimentales pouvant avoir une incidence sur l'efficacité d'incorporation de nucléotides, et fournit un point de référence pour l'interprétation de polypmorphismes. Cette invention concerne également des méthodes d'obtention de populations de moutons résistant à la tremblante.

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