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Diagram illustrating a protocol for site-directed mutagenesis of a plasmid (from Forloni et al. 2018; Cold Spring Harb Protoc doi: 10.1101/pdb.prot097766). The protocol involves using polymerase chain reaction (PCR) with primers containing the desired mutation to amplify a mutated plasmid; this is followed by digestion of the original plasmid with a methylation-sensitive restriction enzyme. In this issue, Narendra Wajapeyee and colleagues introduce methods such as this one for in vitro mutagenesis (doi: 10.1101/pdb.top097733).