C - Chemistry – Metallurgy – 12 – P
Patent
C - Chemistry, Metallurgy
12
P
C12P 19/34 (2006.01) C07H 21/02 (2006.01) C12N 15/10 (2006.01)
Patent
CA 2454319
The present invention provides compositions and improved methods for multi- site directed mutagenesis and DNA shuffling. The present compositions and methods provide increased mutation frequency and increase number of transformants which allow one to sequence only a few clones in order to identify the correct mutants and to obtain the desired mutant by screening large number of transformants in a short time. Moreover, the inclusion of FEN- 1,, PEF and optimized buffer and cycling conditions provided in the present invention should also facilitate random mutagenized library construction and the mutagenesis of large or difficult templates.
La présente invention concerne des compositions et des méthodes améliorées de mutagenèse dirigée contre plusieurs sites et de réarrangement d'ADN. Les compositions et les méthodes de l'invention présentent une fréquence de mutation accrue et un nombre accru de transformants qui ne permettent de séquencer que quelques clones afin d'identifier les mutants appropriés et d'obtenir le mutant désiré par criblage d'un grand nombre de transformants dans un bref laps de temps. De plus, l'inclusion de FEN-1, de PEF, et de conditions optimisées des cycles et des tampons, devrait également faciliter la construction aléatoire d'une bibliothèque mutagénisée et la mutagenèse de grandes matrices ou de matrices difficiles.
Cline Janice M.
Hogrefe Holly H.
Stratagene
Torys Llp
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