Site-directed mutagenesis

Site-directed mutagenesis is a laboratory method for introducing a deliberately chosen change at a specific DNA sequence position. It enables researchers to test how particular genetic changes affect a gene or its product.

Mutagenic primer: A short DNA oligonucleotide designed to introduce a chosen sequence change during DNA synthesis. Its altered bases encode the intended mutation in many primer-based protocols.
DNA: A nucleic acid that stores hereditary information in most organisms. The method alters DNA sequence directly, usually in a cloned target.
Alanine scanning: A mutational strategy that replaces selected protein residues with alanine to probe their contributions. It uses targeted substitutions to identify residues important for a protein's activity.
Random mutagenesis: A strategy that creates mutations at unpredictable positions in genetic material. Unlike targeted mutagenesis, it explores changes without specifying each altered site.
Polymerase chain reaction: A technique that amplifies selected DNA sequences through repeated cycles of heating and DNA synthesis. PCR-based protocols copy a template while incorporating mutation-bearing primers.
Gene: A DNA sequence that contributes to a functional product, such as an RNA or protein. Mutations are introduced into genes to test their roles or alter their products.
Structure–function relationship: The connection between a molecule's structural features and its biological activity. Targeted mutations test whether specific sequence features determine function.
Saturation mutagenesis: A strategy that systematically substitutes many or all possible amino acids at selected positions. It expands a chosen site into a library rather than testing only one designed change.
DNA polymerase: An enzyme that synthesizes DNA by adding nucleotides to a growing strand using a template. The polymerase copies the target region and can carry the primer-encoded change into new DNA.
Nucleotide: A molecular unit of DNA or RNA composed of a sugar, phosphate, and nitrogenous base. A chosen mutation changes one or more nucleotide identities or positions.

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