Circular polymerase extension cloning of complex gene libraries and pathways.

Published

Journal Article

High-throughput genomics 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 the development of a novel and extremely simple cloning method, circular polymerase extension cloning (CPEC). 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. In this study, we elucidate the CPEC reaction mechanism and demonstrate its usage in demanding synthetic biology applications such as one-step assembly and cloning of complex combinatorial libraries and multi-component pathways.

Full Text

Cited Authors

  • Quan, J; Tian, J

Published Date

  • January 2009

Published In

Volume / Issue

  • 4 / 7

Start / End Page

  • e6441 -

PubMed ID

  • 19649325

Pubmed Central ID

  • 19649325

Electronic International Standard Serial Number (EISSN)

  • 1932-6203

International Standard Serial Number (ISSN)

  • 1932-6203

Digital Object Identifier (DOI)

  • 10.1371/journal.pone.0006441

Language

  • eng