Mutation Detection


Mutation detection is essential for the practice of modern biology. Because sequencing deoxyribonucleic acid (DNA) to detect changes in DNA (mutations) is not cost‐effective, a range of methods have been developed to detect mutations for the first time and a different set of methods to look for previously defined mutations.

Keywords: genotyping; mutation discovery; heteroduplex; point mutations; single nucleotide polymorphism

Figure 1.

Heteroduplex formation producing mismatched base pairs is essential for many mutation detection methods. Hybridization of mutant and wild‐type sequences is shown to produce two complementary heteroduplexes, and the original homoduplexes reform. Full lines represent the DNA sequence.

Figure 2.

Schematic representations of methods to detect unknown mutations: (a) ribonuclease; (b) denaturing gradient gel electrophoresis; (c) single‐strand conformation polymorphism; (d) heteroduplex; (e) enzyme and chemical cleavage of mismatch; (f) protein truncation test. Full lines represent the DNA sequence (except where RNA is indicated by R). Mutations are represented by an X. Arrows represent cleavage.

Figure 3.

Schematic representation of methods to detect known mutations: (a) restriction enzyme cleavage; (b) allele‐specific oligonucleotide; (c) primer extension; (d) allele‐specific amplification; (e) oligonucleotide ligation; (f) Taqman. Full lines represent the DNA sequence. Mutations are represented by an X. L and F represent labels, usually fluorescent, and Q, a quencher. Diagonal arrow represent cleavage.


Further Reading

Antonarakis SE and the Nomenclature Working Group (1998) Recommendations for a nomenclature system for human gene mutations. Human Mutation 11: 1–3.

Cotton RGH (1997) Mutation Detection. Oxford, UK: Oxford University Press.

Dracopoli NC, Haines JL, Korf BR, et al. (1996) (eds.) Current Protocols in Human Genetics. New York, NY: Wiley Liss.

Eng C and Vijg J (1997) Genetic testing: the problems and promise. Nature Biotechnology 15: 422–426.

Kwok P‐Y (2000) High‐throughput genotyping assay approaches. Pharmacogenomics 1: 1–6.

Kwok P‐Y (ed.) (2002) The Nucleic Acid Protocols Handbook. Totowa, NJ: Humana Press.

Semino O, Passarino G, Oefner PJ, et al. (2000) The genetic legacy of Paleolithic Homo sapiens sapiens in extant Europeans: a Y chromosome perspective. Science 290: 1155–1158.

Syvanen A‐C (2001) Nature Reviews Genetics 2: 930–942.

Weaver TA (2000) High‐throughput SNP discovery and typing for genome‐wide genetic analysis. New Technologies for Life Sciences: A Trends Guide, pp. 36–42. Amsterdam: Elsevier Science Limited.

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Cotton, RGH(Jan 2006) Mutation Detection. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1038/npg.els.0005672]