Abstract
BackgroundThe subcloning of a DNA fragment from an entry vector into a destination vector is a routinely performed task in molecular biology labs.ResultsWe 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.ConclusionThe 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.
Highlights
The subcloning of a DNA fragment from an entry vector into a destination vector is a routinely performed task in molecular biology labs
The method relies on a 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
Summary
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 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
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