PCR mutagenesis is a method for generating site-directed mutagenesis

Site-directed mutagenesis

Site-directed mutagenesis, also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, is a molecular biology technique in which a mutation is created at a defined site in a DNA molecule. In general, this form of mutagenesis requires that the wild type gene sequence be known...

. This method can generate mutations (base substitutions, insertions, and deletions) without the need for subcloning into M13-based bacteriophage

Bacteriophage

A bacteriophage is any one of a number of viruses that infect bacteria. They do this by injecting genetic material, which they carry enclosed in an outer protein capsid...

 vectors and for ssDNA rescue. The procedure involves a PCR reaction using a plasmid or chromosomal DNA as template and synthetic oligonucleotide primers containing the desired mutation with each complementary to the opposite strands of the vector.