Cloning PCR products by addition of restriction sites to the termini of amplified DNA.

INTRODUCTION Pairs of oligonucleotide primers used in PCR are often designed with restriction sites in their 5' regions. In many cases, the sites are different in the two primers. In this case, amplification generates a target fragment whose termini now carry new restriction sites that can be used for directional cloning into plasmid vectors. The purified fragment and the vector are digested with the appropriate restriction enzymes, ligated together, and transformed into E. coli.