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.