Plasmid Construction by SLIC or Sequence and Ligation-Independent Cloning

  • Ryan E. Hill
  • Julian J. Eaton-Rye
Part of the Methods in Molecular Biology book series (MIMB, volume 1116)


Sequence and ligation-independent cloning (Nat Methods 4:251–256, 2007) is a powerful tool for the construction of multi-fragment complex plasmids in a simple and efficient manner. Plasmids consisting of 6–7 DNA fragments can be assembled in a single day, with additional 2 days for screening and extraction. SLIC requires PCR products with overlapping regions of 30–40 bp at the 5′ and 3′ ends, T4 DNA polymerase, and an optional RecA protein for construction.

Key words

Synthetic biology SLIC Plasmid construction Molecular biology Cloning 



REH was supported by a Todd Foundation Postgraduate Scholarship in Energy Research and JER acknowledges the support of New Zealand Marsden Contract 08-UOO-043.


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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Ryan E. Hill
    • 1
  • Julian J. Eaton-Rye
    • 1
  1. 1.Department of BiochemistryUniversity of OtagoDunedinNew Zealand

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