Gao Song, Li Yanling, Zhang Jiannan, Chen Hongman, Ren Daming, Zhang Lijun, An Yingfeng
College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
College of Biosciences and Biotechnology, Shenyang Agricultural University, Shenyang 110161, China.
Anal Biochem. 2014 May 15;453:55-7. doi: 10.1016/j.ab.2014.02.031. Epub 2014 Mar 5.
Here we describe a modified version of the digestion-ligation approach for efficient molecular cloning. In comparison with the original method, the modified method has the additional steps of gel purification and a second ligation after the first ligation of the linearized vector and DNA insert. During this process, the efficiency and reproducibility could be significantly improved for both stick-end cloning and blunt-end cloning. As an improvement of the very important molecular cloning technique, this method may find a wide range of applications in bioscience and biotechnology.
在此，我们描述了一种用于高效分子克隆的消化-连接方法的改进版本。与原始方法相比，改进后的方法在将线性化载体和DNA插入片段进行首次连接后，增加了凝胶纯化和第二次连接的步骤。在此过程中，粘端克隆和平端克隆的效率和可重复性都能得到显著提高。作为对这一非常重要的分子克隆技术的改进，该方法可能在生物科学和生物技术领域有广泛的应用。
