| Title | MINIREVIEW] Modern and Simple Construction of Plasmid: Saving Time and Cost |
|---|---|
| Author | Hideki Nakayama and Nobuo Shimamoto* |
| Address | Faculty of Life Sciences, Kyoto Sangyo University, Kamigamo-Motoyama, Kita-Ku, Kyoto, 603-8555, Japan |
| Bibliography | Journal of Microbiology, 52(11),891-897, 2014, |
| DOI | 10.1007/s12275-014-4501-6 |
| Key Words | plasmid, cloning, recombineering, PCR cloning, fusion PCR, Gibson assembly, SLiCE |
| Abstract | Construction of plasmids has been occupying a significant fraction of laboratory work in most fields of experimental biology. Tremendous effort was made to improve the traditional method for constructing plasmids, in which DNA fragments digested with restriction enzymes were ligated. However, the traditional method remained to be a standard protocol more than 40 years. At last, several recent inventions are rapidly and completely replacing the traditional method, because they are far quicker with less cost, and requiring less material. We here introduce three such methods that cover up most of the cases. Moreover, they are complementary with each other. Our lab protocols are provided for “no strain, no pain” construction of plasmids. |