Subcloning of nucleic acid fragments
The invention deals with subcloning of a nucleic acid fragment from a source vector into any target vector. It can be carried out in one step; the result is available within a few hours, easy to handle, independent from the type and sequences of the target vector, and independent from resistance markers in the target vector.
This procedure facilitates one of the most frequent routine tasks in molecular biology: the subcloning of nucleic acid fragments from one vector into another. As the only precondition for this, a source vector with two sequence sections (3) is required, both of which carry a recognition site for an outside cutter restriction enzyme and bordering nucleotides which form a specific base overlap with the outside cutter restriction enzyme during cleavage. The invention makes use of the fact that outside cutter restriction enzymes recognise a recognition site of 4-8 base pairs, while the cleavage site is located within a defined distance from the recognition site.
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