title: One-pot Simple methodology for CAssette Randomization and Recombination for focused directed evolution (OSCARR).
abstract: The OSCARR methodology (One-pot Simple methodology for CAssette Randomization and Recombination) bridges the gap between site-directed mutagenesis and full randomization by making use of carefully designed mutagenic cassettes and an optimized one-pot megaprimer PCR. The method is especially suited to construct libraries of up to ten randomized codons for focused directed evolution, exhibits up to 97 % efficiency in the amplification of mutated over wild-type products, and is sufficiently versatile to allow mutagenesis and recombination of several cassettes within the same gene.
authors: Center for Molecular Biology, UAM-CSIC, Madrid, Spain.
authors: Aurelio
authors: A
authors: Hidalgo
authors: Anna
authors: A
authors: Schließmann
authors: Uwe T
authors: UT
authors: Bornscheuer
mesh_terms: D019020:Directed Molecular Evolution / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction