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

ABSTRACT:

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 enz

SUBMITTER: Fromme T 

PROVIDER: S-EPMC2241830 | biostudies-literature | 2007 Nov

REPOSITORIES: biostudies-literature