A modified version of the digestion-ligation cloning method for more efficient molecular cloning.
A modified version of the digestion-ligation cloning method for more efficient molecular cloning.
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DOI：
10.1016/j.ab.2014.02.031
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发表时间：
2014-05
影响因子：
2.9
通讯作者：
Song Gao;Yanling Li;Jiannan Zhang;Hongman Chen;Daming Ren;Lijun Zhang;Y. An
中科院分区：
文献类型：
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作者：
Song Gao;Yanling Li;Jiannan Zhang;Hongman Chen;Daming Ren;Lijun Zhang;Y. An
在这里，我们描述了一种改进的消化-连接方法，用于高效的分子克隆。与原方法相比，改进的方法增加了凝胶纯化步骤和线性化载体和DNA插入片段第一次连接后的第二次连接步骤。在此过程中，杆端克隆和钝端克隆的效率和重复性都可以显著提高。作为一种非常重要的分子克隆技术的改进，该方法在生物科学和生物技术中具有广泛的应用前景。
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.