Rapid single step subcloning procedure by combined action of type II and type IIs endonucleases with ligase

被引：9

作者：

Fromme T. ^{[1
]}

Klingenspor M. ^{[1
]}

机构：

[1] Department of Animal Physiology, Faculty of Biology, Philipps-University

来源：

Journal of Biological Engineering | / 1卷 / 1期

关键词：

Ligase; Destination Vector; Entry Vector; BglII Site; Restriction Enzyme Recognition Site;

D O I：

10.1186/1754-1611-1-7

中图分类号：

学科分类号：

摘要：

Background: The subcloning of a DNA fragment from an entry vector into a destination vector is a routinely performed task in molecular biology labs. Results: 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. Conclusion: 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. © 2007 Fromme and Klingenspor; licensee BioMed Central Ltd.

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