Journal
GENETICS AND MOLECULAR RESEARCH
Volume 14, Issue 2, Pages 3466-3473Publisher
FUNPEC-EDITORA
DOI: 10.4238/2015.April.15.10
Keywords
DpnI digestion; High-fidelity DNA polymerase; Site-directed mutagenesis
Funding
- State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences [SKLVEB2011KFKT003]
Ask authors/readers for more resources
Gene mutation plays an important role in molecular biological studies. A highly efficient one-step polymerase chain reaction-based mutagenesis technique for site-directed mutagenesis was developed in this study. One complementary pair of primers was designed that contained the desired mutations in the middle of the primers. The amplification products of mutation were amplified using a high-fidelity DNA polymerase and the original plasmid templates were digested by DpnI. This method was successfully used to introduce mutations in two different-sized plasmids (12 and 6 kb) with high efficiency. The results indicate that this technique can be widely used to introduce any plasmid mutations quickly and efficiently.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available