Dna joining method

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 15/66 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2402321

The present invention provides a method to directionally clone any linear template DNA molecule into any linearized vector. The vector ends may be generated from any restriction enzyme cleavage. The method does not require a ligation step nor the use of carefully controlled conditions as is required with methods involving specific exonucleases alone. It has been determined that specific DNA polymerases are able to efficiently join one or more linear DNA molecules sharing ends with appropriate complementation.

L'invention concerne un procédé de clonage directionnel d'une quelconque molécule linéaire d'ADN modèle, pour obtenir un quelconque vecteur linéarisé. Les extrémités du vecteur peuvent être produites à partir d'un quelconque clivage par une enzyme de restriction. Ce procédé ne nécessite pas d'étape de ligature ni de conditions d'exécution strictes, comme c'est le cas dans des procédés impliquant l'utilisation d'exonucléases spécifiques seules. Il a été déterminé que des ADN polymérases spécifiques sont capables d'assembler de manière efficace une ou plusieurs molécules linéaires d'ADN partageant des extrémités possédant une complémentarité adéquate.

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