In this method, a fragment of DNA is synthesized, and then inserted into a plasmid. It involves the cleavage by a restriction enzyme at a site in the plasmid and subsequent ligation of a pair of complementary oligonucleotides containing the mutation in the gene of interest to the plasmid.

What are the advantages of site-directed mutagenesis?

Advantages. Both primers at the mutation site contain the desired mutation. Short and easy PCR reactions. Ideal for introducing multiple mutations (close or distant).

What are the types of site-directed mutagenesis?

Depending on the number of sites to be mutated, site‐directed mutagenesis can be divided into two types: simple or multiple mutations [2]. For single mutations, methods are based on the amplification of double‐stranded DNA from plasmids using complementary oligonucleotides carrying the mutation of interest [3].