Revised Mechanism and Improved Efficiency of the QuikChange Site-Directed Mutagenesis Method

Yongzhen Xia; Luying Xun

doi:10.1007/978-1-4939-6472-7_25

Revised Mechanism and Improved Efficiency of the QuikChange Site-Directed Mutagenesis Method

Methods Mol Biol. 2017:1498:367-374. doi: 10.1007/978-1-4939-6472-7_25.

Authors

Yongzhen Xia¹, Luying Xun²

Affiliations

¹ State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, China.
² School of Molecular Biosciences, Washington State University, Pullman, WA, 99164-7520, USA.

PMID: 27709589
DOI: 10.1007/978-1-4939-6472-7_25

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.

Keywords: Overlap PCR; PCR; QuikChange; Site-directed mutagenesis.

MeSH terms

Base Sequence / genetics
DNA Primers / genetics
Mutagenesis / genetics*
Mutagenesis, Site-Directed / methods*
Polymerase Chain Reaction / methods

Substances

DNA Primers