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Single-Strand Conformational Polymorphism Analysis: Basic Principles and Routine Practice
Abstract
Single-strand conformational polymorphism (SSCP) analysis is a simple and sensitive technique for mutation detection and genotyping. The principle of SSCP analysis is based on the fact that single-stranded DNA has a defined conformation. Altered conformation due to a single base change in the sequence can cause single-stranded DNA to migrate differently under nondenaturing electrophoresis conditions. Therefore wild-type and mutant DNA samples display different band patterns. SSCP analysis involves the following four steps: (1) polymerase chain reaction (PCR) amplification of DNA sequence of interest; (2) denaturation of double-stranded PCR products; (3) cooling of the denatured DNA (single-stranded) to maximize self-annealing; (4) detection of mobility difference of the single-stranded DNAs by electrophoresis under non-denaturing conditions. Several methods have been developed to visualize the SSCP mobility shifts. These include the incorporation of radioisotope labeling, silver staining, fluorescent dye-labeled PCR primers, and more recently, capillary-based electrophoresis. Silver staining is simple, rapid, and cost-effective, and can be routinely performed in clinical laboratories. The use of SSCP analysis to discover and genotype single-nucleotide polymorphisms (SNPs) has been widely applied to the genetics of hypertension, including both monogenic (e.g., Liddle's syndrome) and polygenic disorders (e.g., essential hypertension).
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Altered conformation due to a single base change in the DNA can cause different band patterns on non-denaturing gels which separate mutant and wild-type DNA.
“Altered conformation due to a single base change in the DNA can cause different band patterns on non-denaturing gels which separate mutant and wild-type DNA (Zhu, 2005).”
From Genetic resources and precise gene editing for targeted improvement of barley abiotic stress tolerance (2023) · cites this paper as Zhu, 2005
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Schreiber, L., Nader-Nieto, A. C., Schönhals, E. M., Walkemeier, B., & Gebhardt, C. (2014) · G3 Genes|Genomes|Genetics · doi:10.1534/g3.114.012377 · Cited by 81
Karunarathne, S., Walker, E., Sharma, D., Li, C., & Han, Y. (2023) · Journal of Zhejiang University-SCIENCE B · doi:10.1631/jzus.b2200552 · Cited by 18
“Altered conformation due to a single base change in the DNA can cause different band patterns on non-denaturing gels which separate mutant and wild-type DNA (Zhu, 2005).”
Influential
Metadata
- Authors
- Dong, Y., & Zhu, H.
- Affiliations
- Augusta University (US)
- Venue
- Hypertension
- Publisher
- Humana Press
- Published
- 2005-07-21
- Volume
- 108
- Pages
- 149-158
- OpenAlex
- W133571668
- Cited by
- 44
- References
- 24
- Citations by year
- 2025: 3 · 2024: 3 · 2023: 5 · 2022: 1 · 2021: 2 · 2020: 5 · 2019: 5 · 2018: 1 · 2017: 5 · 2016: 2 · 2014: 2 · 2013: 1
- FWCI
- 5.99
- Type
- book chapter
- Language
- en
- Open access
- closed
- Locations
- Humana Press eBooks (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genetic Associations and Epidemiology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Genomics and Rare Diseases
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Keywords
- single-strand conformation polymorphism, mutation detection, genotyping, single nucleotide polymorphisms, silver staining, self-annealing, capillary electrophoresis, hypertension genetics, Liddle syndrome, essential hypertension
- MeSH terms
- DNA, Electrophoresis, Polymorphism, Single-Stranded Conformational
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