Circular polymerase extension cloning.

ABSTRACT

High-throughput genomics, proteomics, and the emerging field of synthetic biology demand ever more convenient, economical, and efficient technologies to assemble and clone genes, gene libraries, and synthetic pathways. Here, we describe an extremely simple, efficient, and cost-effective cloning method, circular polymerase extension cloning (CPEC), for complex, combinatorial, or multi-fragment assembly as well as routine cloning. This method uses a single polymerase to assemble and clone multiple inserts with any vector in a one-step reaction in vitro. No restriction digestion, ligation, or single-stranded homologous recombination is required.