IVA cloning: A single-tube universal cloning system exploiting bacterial In Vivo Assembly.

ABSTRACT: In vivo homologous recombination holds the potential for optimal molecular cloning, however, current strategies require specialised bacterial strains or laborious protocols. Here, we exploit a recA-independent recombination pathway, present in widespread laboratory E.coli strains, to develop IVA (In Vivo Assembly) cloning. This system eliminates the need for enzymatic assembly and reduces all molecular cloning procedures to a single-tube, single-step PCR, performed in <2 hours from setup to transformation. Unlike other methods, IVA is a complete system, and offers significant advantages over alternative methods for all cloning procedures (insertions, deletions, site-directed mutagenesis and sub-cloning). Significantly, IVA allows unprecedented simplification of complex cloning procedures:

SUBMITTER: Garcia-Nafria J 

PROVIDER: S-EPMC4893743 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature