DNA Cloning and Assembly Methods pp 103-117

Part of the Methods in Molecular Biology book series (MIMB, volume 1116) | Cite as

Circular Polymerase Extension Cloning

  • Jiayuan Quan
  • Jingdong Tian
Protocol

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.

Key words

Molecular cloning Genetic assembly Library cloning DNA polymerase 

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Jiayuan Quan
    • 1
    • 2
  • Jingdong Tian
    • 1
    • 2
  1. 1.Department of Biomedical Engineering and the Institute for Genome Sciences and PolicyDuke UniversityDurhamUSA
  2. 2.Tianjin Institute of Industrial BiotechnologyChinese Academy of SciencesTianjinChina

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