Rapid analysis of variations in a genome

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01)

Patent

CA 2477611

The invention provides a method useful for determining the sequence of large numbers of loci of interest on a single or multiple chromosomes. The method utilizes an oligonucleotide primer that contains a recognition site for a restriction enzyme such that digestion with the restriction enzyme generates a 5' overhang containing the locus of interest. The 5' overhang is used as a template to incorporate nucleotides, which can be detected. The method is especially amenable to the analysis of large numbers of sequences, such as single nucleotide polymorphisms, from one sample of nucleic acid.

Cette invention concerne une méthode utile pour déterminer la séquence d'un grand nombre de locus d'intérêt sur un chromosome simple ou des chromosomes multiples. La méthode de l'invention met en oeuvre une amorce oligonucléotidique qui contient un site de reconnaissance pour une enzyme de restriction, tel que la digestion avec l'enzyme de restriction génère une extrémité 5' protubérante contenant le locus d'intérêt. L'extrémité 5' protubérante est utilisée comme modèle pour incorporer des nucléotides pouvant être détectés. Cette méthode convient particulièrement pour l'analyse d'un grand nombre de séquences, telles que des polymorphismes nucléotidiques simples issus d'un échantillon d'acide nucléique.

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