Department of Animal Physiology, Faculty of Biology, Philipps-University, D-35043 Marburg, Germany.
J Biol Eng. 2007 Nov 26;1:7. doi: 10.1186/1754-1611-1-7.
The subcloning of a DNA fragment from an entry vector into a destination vector is a routinely performed task in molecular biology labs.
We here present a novel benchtop procedure to achieve rapid recombination into any destination vector of choice with the sole requirement of an endonuclease recognition site. The method relies on a specifically designed entry vector and the combined action of type II and type IIs endonucleases with ligase. The formulation leads to accumulation of a single stable cloning product representing the desired insert carrying destination vector.
The described method provides a fast single step procedure for routine subcloning from an entry vector into a series of destination vectors with the same restriction enzyme recognition site.
从入口载体中将 DNA 片段亚克隆到目的载体中是分子生物学实验室中常规进行的任务。
我们在这里提出了一种新颖的台式程序，可仅通过内切酶识别位点将其快速重组到任何所需的目的载体中。该方法依赖于专门设计的入口载体以及 II 型和 II 型内切酶与连接酶的联合作用。该配方导致积累了一个单一的稳定克隆产物，代表带有目的载体的所需插入物。
所描述的方法提供了一种快速的单步程序，可用于从入口载体常规亚克隆到具有相同限制酶识别位点的一系列目的载体中。
