A Sequence- and Ligation-Independent Cloning (SLIC) Procedure for the Insertion of Genes into a Plasmid Vector

Molecular cloning is a routine technique for many laboratories with applications from genetic engineering to recombinant protein expression. While restriction-ligation cloning can be slow and inefficient, ligation-independent cloning uses long single-stranded overhangs generated by T4 DNA polymerase's 3' exonuclease activity to anneal the insert and plasmid vector prior to transformation. This chapter describes a fast, high-efficiency protocol for inserting one or more genes into a vector using sequence- and ligation-independent cloning (SLIC).