Circular polymerase extension cloning for high-throughput cloning of complex and combinatorial DNA libraries.

Published

Journal Article

High-throughput genomics, proteomics and synthetic biology studies require ever more efficient and economical strategies to clone complex DNA libraries or variants of biological modules. In this paper, we provide a protocol for a sequence-independent approach for cloning complex individual or combinatorial DNA libraries, and routine or high-throughput cloning of single or multiple DNA fragments. The strategy, called circular polymerase extension cloning (CPEC), is based on polymerase overlap extension and is therefore free of restriction digestion, ligation or single-stranded homologous recombination. CPEC is highly efficient, accurate and user friendly. Once the inserts and the linear vector have been prepared, the CPEC reaction can be completed in 10 min to 3 h, depending on the complexity of the gene libraries.

Full Text

Cited Authors

  • Quan, J; Tian, J

Published Date

  • February 3, 2011

Published In

Volume / Issue

  • 6 / 2

Start / End Page

  • 242 - 251

PubMed ID

  • 21293463

Pubmed Central ID

  • 21293463

Electronic International Standard Serial Number (EISSN)

  • 1750-2799

International Standard Serial Number (ISSN)

  • 1754-2189

Digital Object Identifier (DOI)

  • 10.1038/nprot.2010.181

Language

  • eng