Method of mapping restriction sites in polynucleotides

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2295325

The invention provides a method for constructing a high resolution physical map of a polynucleotide. In accordance with the invention, the polynucleotide is digested successively with at least two different restriction endonucleases and the ends of the restriction fragments are sequenced after each digestion. In this manner, restriction fragments having sequenced ends are produced that can be aligned by their sequences to give a physical map of the polynucleotide. Preferably, restriction fragment ends are sequenced by massively parallel signature sequencing (MPSS), or a like parallel sequencing technique.

La présente invention concerne un procédé d'établissement d'une cartographie physique haute résolution d'un polynucléotide. Selon cette invention, le polynucléotide est digéré successivement avec au moins deux endonucléases de restriction différentes, les extrémités des fragments de restriction étant séquencées après chaque digestion. Ainsi, les fragments de restriction pourvus d'extrémités séquencées sont obtenus et peuvent être alignés par leurs séquences de manière à obtenir une cartographie physique du nucléotide. De préférence, les extrémités des fragments de restriction sont séquencées par un procédé de séquençage massif de signatures parallèles, ou par une technique de séquençage parallèle similaire.

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