DNA Cloning and Assembly Methods pp 25-36

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

| Cite as

Plasmid Construction by SLIC or Sequence and Ligation-Independent Cloning

  • Ryan E. Hill
  • Julian J. Eaton-Rye
Protocol

Abstract

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 

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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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