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Site-Directed Mutagenesis (SDM) allows for changes in the DNA sequence of plasmids using polymerase chain reaction (PCR). It is a reliable, accessible, and rapid method which is the common initial step of many biochemial or genetic experiments. Here we describe the various different forms of SDM before giving a detailed method for the introduction of substitutions, insertions, or deletions using a fast, ligation-free protocol, followed by colony PCR to screen for mutated sequences.

Original publication




Journal article


Methods Mol Biol

Publication Date





87 - 95


Colony PCR, Genetic alteration, Mutation, Plasmid manipulation, Polymerase chain reaction, Site-directed mutagenesis, Mutagenesis, Site-Directed, Polymerase Chain Reaction