title: Long distance multiple-site directed plasmid mutagenesis by one-step PCR using non-overlapped primers.
abstract: Site-directed mutagenesis is a very important technique in molecular biological researches. We have developed a new method for long distance multiple-site plasmid mutation by one-step PCR using non-overlap primers. These primers were carefully designed and contained desired mutations in the middle of the primers flanked with 18-25 bp of correct sequence. One pair of the primers was able to generate a short megaprimer. Decreases in the concentrations of these primers increased efficiency of the multiple-site plasmid mutation. All of the mutant PCRs were performed at a common annealing temperature at 55 °C. This method could be widely used in all multiple-site plasmid mutations.
authors: Key Laboratory of Brain Functional Genomics, Ministry of Education, Shanghai Key Laboratory of Brain Functional Genomics, East China Normal University, Shanghai, 200062, People's Republic of China.
authors: Yuling
authors: Y
authors: Bu
authors: Hui
authors: H
authors: Wang
authors: Jing
authors: J
authors: Li
authors: Zhaoxin
authors: Z
authors: Zhang
authors: Yinghe
authors: Y
authors: Hu
authors: Zhiliang
authors: Z
authors: Xu
mesh_terms: D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D013696:Temperature
chemical_list: D017931:DNA Primers
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