期刊
GENETICS AND MOLECULAR RESEARCH
卷 14, 期 2, 页码 3466-3473出版社
FUNPEC-EDITORA
DOI: 10.4238/2015.April.15.10
关键词
DpnI digestion; High-fidelity DNA polymerase; Site-directed mutagenesis
资金
- State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences [SKLVEB2011KFKT003]
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.
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