Figure 3: Schematic diagram of mutagenesis by quick change. In the first step of mutagenesis by an overlap extension using mutagenic complementary primers, both containing the desired mutation, a mutated plasmid containing staggered nicks are generated. In the second step by digestion with the restriction enzyme DpnI the methylated parental DNA template is removed. In the third step the nicked mutated vector is transformed into an E. coli recAstrain.