Chain reaction cloning

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 19/34 (2006.01) A61K 39/21 (2006.01) A61K 39/245 (2006.01) C07K 14/035 (2006.01) C07K 14/16 (2006.01) C12N 15/10 (2006.01) C12N 15/52 (2006.01) A61K 48/00 (2006.01)

Patent

CA 2253880

Chain reaction cloning methods and reagents and kits for performing such methods are disclosed. Chain reaction cloning methods provide a convenient one step process that will allow site-specific ligation of DNA molecules by thermostable DNA ligases. A number of applications are disclosed for chain reaction cloning including site-specific ligation of DNA fragments generated by restriction enzyme digestion, DNAse digestion, chemical cleavage, enzymatic or chemical synthesis, and PCR amplification.

L'invention concerne des procédés de clonage, des réactifs et des trousses permettant de mettre en oeuvre les procédés proposés. Les procédés de clonage par réaction en chaîne permettent de réduire le processus à une seule étape pendant laquelle s'opère la ligature dirigée des molécules de l'ADN par des ADN ligases thermostables. On propose plusieurs applications du clonage par réaction en chaîne, qui incluent la ligature dirigée des fragments de l'ADN grâce à la digestion par des enzymes de restriction ou par l'ADN-ase, au clivage chimique, à la synthèse enzymatique ou chimique et à l'amplification de la réaction en chaîne de la polymérase (PCR).

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