An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy.

ABSTRACT:

Background

Widely used restriction-dependent cloning methods are labour-intensive and time-consuming, while several types of ligase-independent cloning approaches have inherent limitations. A rapid and reliable method of cloning native DNA sequences into desired plasmids are highly desired.

Results

This paper introduces ABI-REC, a novel strategy combining asymmetric bridge PCR with intramolecular homologous recombination in bacteria for native DNA cloning. ABI-REC was developed to precisely clone inserts into defined location in a directional manner within recipient plasmids. It featured an asymmetric 3-primer PCR performed in a single tube that could robustly amplify a chimeric insert-plasmid DNA sequence with homologous arms at both ends. Intramolecular homologous recombin

SUBMITTER: Bi Y 

PROVIDER: S-EPMC3408372 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature