T4 DNA polymerase improves the efficiency of multiple site-directed mutagenesis

Abstract

Site-directed mutagenesis (SDM) is a useful tool to study the functions of regulatory sequences of DNA and RNA, and the structures and functions of proteins. Numerous methods have been developed for either single or multiple SDM (MSDM). However, MSDM is sometimes difficult. Here we demonstrated that T4 DNA polymerase greatly enhanced the efficiency of MSDM. Moreover, we have also showed that it is efficient to clone multiple specific mutation-containing sequences simultaneously.

Keywords

• Multiple site-directed mutagenesis
• complementary annealing mediated by exonuclease
• T4 DNA polymerase
• DNA assembly