Generation of Multiple Site-Specific Mutations by Polymerase Chain Reaction

Generating mutant proteins has become a routine strategy for molecular and structural biologists, to understand the structure-function relationship of a given protein of interest. In this, site-directed mutagenesis has proven to be a valuable technique. After the advent of polymerase chain reaction (PCR), PCR- mediated mutagenesis has been the method of choice to change from one amino acid to another, which has replaced the classic Kunkel’s method (1 ).