Tripp Valerie T, Maza Johnathan C, Young Douglas D
Department of Chemistry, The College of William & Mary, Williamsburg, VA 23187 USA.
J Chem Biol. 2013 May 21;6(3):135-40. doi: 10.1007/s12154-013-0095-4.
The introduction of exogenous DNA into Escherichia coli is a cornerstone of molecular biology. Herein, we investigate two new mechanisms for bacterial transformation involving either the use of microwave irradiation or a freeze-thaw protocol in liquid nitrogen. Ultimately, both methods afforded successful transfer of plasmid DNA into bacterial cells, with the freeze-thaw technique yielding efficiencies of ~10(5). More importantly, both techniques effectively eliminated the need for the preparation of competent cells.
将外源DNA导入大肠杆菌是分子生物学的基石。在此,我们研究了两种细菌转化的新机制,一种涉及使用微波辐射,另一种涉及在液氮中的冻融方案。最终,两种方法都成功地将质粒DNA转移到细菌细胞中,冻融技术产生的效率约为10⁵。更重要的是,这两种技术都有效地消除了制备感受态细胞的必要性。