title: Revised Mechanism and Improved Efficiency of the QuikChange Site-Directed Mutagenesis Method.
abstract: Site-directed mutagenesis has been widely used for the substitution, addition or deletion of nucleotide residues in a defined DNA sequence. QuikChange™ site-directed mutagenesis and its related protocols have been widely used for this purpose because of convenience and efficiency. We have recently demonstrated that the mechanism of the QuikChange™ site-directed mutagenesis process is different from that being proposed. The new mechanism promotes the use of partially overlapping primers and commercial PCR enzymes for efficient PCR and mutagenesis.
authors.affiliation: State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, China.
authors.forename: Yongzhen
authors.initials: Y
authors.lastname: Xia
authors.affiliation: School of Molecular Biosciences, Washington State University, Pullman, WA, 99164-7520, USA.
authors.forename: Luying
authors.initials: L
authors.lastname: Xun
mesh_terms: D001483:Base Sequence / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D016296:Mutagenesis / Q000235:genetics*; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods
chemical_list: D017931:DNA Primers
keywords: Overlap PCR; PCR; QuikChange; Site-directed mutagenesis