Overview
Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA. During several rounds of thermal cycling, both strands of the plasmid DNA are amplified in a linear fashion, generating a mutated plasmid containing staggered nicks on opposite strands. By Joseph Sambrook and David W. Russell.
Details
Details of this protocol, In Vitro Mutagenesis Using Double-Stranded DNA Templates: Selection Of Mutants With DpnI (Subscription Required), are located on a web site other than Biocompare Protocols.