Rapid single step subcloning procedure by combined action of type II and type IIs endonucleases with ligase.
Rapid single step subcloning procedure by combined action of type II and type IIs endonucleases with ligase.
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DOI：
10.1186/1754-1611-1-7
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发表时间：
2007-11-26
影响因子：
5.6
通讯作者：
Klingenspor M
中科院分区：
文献类型：
--
作者：
Fromme T;Klingenspor M
从入口载体到目标矢量的DNA片段的亚克隆是分子生物学实验室的常规任务。
我们在这里提出了一种新型的台式程序，可以将快速重组重组到任何选择的目的地向量，唯一需要核酸内切酶识别位点。该方法依赖于专门设计的入口向量以及II型和类型IIS核酸内切酶与连接酶的合并作用。该公式导致单个稳定克隆产物的积累，代表所需的插入携带目标向量。
所述方法为从入口向量进行例行亚克隆提供了一个快速的单个步骤过程，从一个具有相同限制酶识别位点的目标向量进行了一个。
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.