title: A method for multi-site-directed mutagenesis based on homologous recombination.
abstract: We have developed an efficient method for the simultaneous introduction of up to three mutations in a plasmid DNA via homologous recombination. The strategy is compatible with a variety of mutations, including degenerate codons in plasmids of different sizes. In contrast to other methodologies, this approach employs the same set of reagents for both single- and multi-site mutagenesis assays, minimizes the required protocol steps, and exhibits remarkably high mutagenesis efficiencies.
authors.affiliation: Life Technologies, Carlsbad, CA 92008, USA.
authors.forename: Xiquan
authors.initials: X
authors.lastname: Liang
authors.forename: Lansha
authors.initials: L
authors.lastname: Peng
authors.forename: Ke
authors.initials: K
authors.lastname: Li
authors.forename: Todd
authors.initials: T
authors.lastname: Peterson
authors.forename: Federico
authors.initials: F
authors.lastname: Katzen
mesh_terms: D004247:DNA / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli; D059765:Homologous Recombination; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D017931:DNA Primers; D004247:DNA