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.