C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/10 (2006.01) C12N 15/66 (2006.01) C12Q 1/68 (2006.01)
Patent
CA 2227413
DNA is cloned and labeled in a sequence-specific manner. The DNA is digested with one or more restriction enzymes which produce 3' recessed ends. A desired fragment is protected from elongation by DNA polymerase by addition of E. coli RecA protein and oligonucleotides about 30 to 60 bases in length complementary to the 3' recessed ends of the digested fragment. RecA and DNA polymerase are then inactivated, leaving only the desired fragment with 3' recessed ends which is then ligated into a vector containing complementary 3' recessed ends.
De l'ADN est cloné et marqué d'une façon spécifique aux séquences. L'ADN est digéré avec une ou plusieurs enzymes de restriction qui produisent des extrémités échancrées 3'. Un fragment désiré est protégé de l'allongement par l'ADN polymérase par addition de protéine RecA de E. Coli et d'oligonucléotides ayant environ 30 à 60 bases de longueur, complémentaires des extrémités échancrées 3' du fragment digéré. La RecA et l'ADN polymérase sont ensuite rendus inactifs, ce qui laisse seulement le fragment désiré avec des extrémités échancrées 3', qui est ensuite ligaturé dans un vecteur comportant des extrémités échancrées 3' complémentaires.
Camerini-Otero Rafael Daniel
Ferrin Lance Joseph
Depar Tment Of Health And Human Services The Government Of The United States Represented By The Secretary
Department Of Health And Human Services The Government Of The United States Of America Represented By Th
Fetherstonhaugh & Co.
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