Overview
Two oligonucleotide primers are hybridized to the same strand of a denatured double-stranded recombinant plasmid. One primer (the mutagenic primer) introduces the desired mutation into the target sequences, and the second primer carries a mutation that destroys a unique restriction site in the plasmid. Both primers are elongated in a reaction catalyzed by bacteriophage T4 or T7 DNA polymerase. Nicks in the strand of newly synthesized DNA are sealed with bacteriophage T4 DNA ligase. By Joseph Sambrook and David W. Russell.
Details
Details of this protocol, Oligonucleotide-Directed Mutagenesis By Elimination Of A Unique Restriction Site (USE Mutagenesis) (Subscription Required), are located on a web site other than Biocompare Protocols.