Electrophoretic methods for mutation detection and DNA sequencing

W Edward Jr. Highsmith

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Citations (Scopus)

Abstract

Since its development in 1985, polymerase chain reaction (PCR) has revolutionized basic and applied research (1,2). In 1993, Mullis was awarded the Nobel Prize in Chemistry for the development of PCR. With DNA or cDNA as a template, millions of copies of a target sequence are generated during the reaction. Introduction of the thermophilic Thermus aquaticus polymerase increased the specificity of the reaction and made automation and routine use possible (3-5). The ability of PCR to produce multiple copies of a discrete portion of the genome has resulted in its incorporation into techniques used in a wide variety of research and clinical applications. An extraordinary range of clinical applications of PCR have emerged, including diagnosis of inherited disease, human leukocyte antigen (HLA) typing, identity testing, infectious disease diagnosis and management, hematologic disease diagnosis and staging, and susceptibility testing for cancer. The development of technically simple and reliable methods to detect sequence variations in specific genes is becoming more important as the number of genes associated with specific diseases grows. DNA sequencing is considered the gold standard for characterization of specific nucleotide alteration(s) that result in genetic disease. Although sequencing was long considered too cumbersome, expensive, and operator dependent for use in the clinical laboratory, a combination of clinical need and improved technology has brought automated DNA sequencing into routine clinical use. However, even though sequencing technology is now firmly entrenched in the clinical molecular diagnostics laboratory, it is still too expensive and time-consuming for all the laboratory's mutation-detection needs. There are a number of PCR-based mutation-detection strategies that can be used to identify both characterized and uncharacterized mutations and sequence variations.

Original languageEnglish (US)
Title of host publicationMolecular Diagnostics: For the Clinical Laboratorian
PublisherHumana Press
Pages85-109
Number of pages25
ISBN (Print)9781588293565
DOIs
StatePublished - 2005

Fingerprint

DNA Sequence Analysis
Polymerase Chain Reaction
Mutation
Taq Polymerase
Nobel Prize
Technology
Inborn Genetic Diseases
Molecular Pathology
Hematologic Diseases
Automation
Disease Management
HLA Antigens
Research
Genes
Communicable Diseases
Nucleotides
Complementary DNA
Genome
DNA
Neoplasms

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Highsmith, W. E. J. (2005). Electrophoretic methods for mutation detection and DNA sequencing. In Molecular Diagnostics: For the Clinical Laboratorian (pp. 85-109). Humana Press. https://doi.org/10.1385/1-59259-928-1:085

Electrophoretic methods for mutation detection and DNA sequencing. / Highsmith, W Edward Jr.

Molecular Diagnostics: For the Clinical Laboratorian. Humana Press, 2005. p. 85-109.

Research output: Chapter in Book/Report/Conference proceedingChapter

Highsmith, WEJ 2005, Electrophoretic methods for mutation detection and DNA sequencing. in Molecular Diagnostics: For the Clinical Laboratorian. Humana Press, pp. 85-109. https://doi.org/10.1385/1-59259-928-1:085
Highsmith WEJ. Electrophoretic methods for mutation detection and DNA sequencing. In Molecular Diagnostics: For the Clinical Laboratorian. Humana Press. 2005. p. 85-109 https://doi.org/10.1385/1-59259-928-1:085
Highsmith, W Edward Jr. / Electrophoretic methods for mutation detection and DNA sequencing. Molecular Diagnostics: For the Clinical Laboratorian. Humana Press, 2005. pp. 85-109
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