Makovets Svetlana
Institute of Cell Biology, University of Edinburgh, Edinburgh, UK.
Methods Mol Biol. 2013;1054:11-43. doi: 10.1007/978-1-62703-565-1_2.
Presented here is a complete molecular cloning protocol consisting of a number of separate but interconnected methods such as preparation of E. coli competent cells; in vitro DNA digestion and ligation; PCR; DNA agarose gel electrophoresis and gel extraction; and screening transformants by colony PCR, analytical restriction digests, and sequencing. The method is described in a lot of details so that it can be easily followed by those with very little relevant knowledge and skills. It also contains many tips that even experienced researchers may find useful.
这里介绍了一个完整的分子克隆方案，该方案由许多独立但相互关联的方法组成，如制备大肠杆菌感受态细胞；体外DNA消化和连接；PCR；DNA琼脂糖凝胶电泳和凝胶提取；以及通过菌落PCR、分析性限制性酶切消化和测序筛选转化体。该方法描述得非常详细，以便相关知识和技能很少的人也能轻松遵循。它还包含许多即使是经验丰富的研究人员也可能会觉得有用的提示。
