Plasmid Construction by SLIC or Sequence and Ligation-Independent Cloning
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 wordsSynthetic 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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