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PubMed · 15095439

Optimum sample medium for single-nucleotide polymorphism and mutation detection by capillary electrophoresis.

Piotr Kozlowski·Wlodzimierz J Krzyzosiak
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Abstract

Capillary electrophoresis (CE) is a versatile analytical platform widely used for nucleic acids analysis. Its applications in research and diagnostics include scanning and screening for mutations and polymorphisms by such reliable methods as single-strand conformation polymorphism (SSCP), heteroduplex analysis (HA), and combined SSCP/HA. This study, aimed at the further development of these methods, is focused on detailed sample-media characteristics. Factors affecting single-strand conformer stability and DNA intake efficiency were analyzed. The sample media optimal for efficient mutation or SNP detection were determined, and complex SSCP-CE patterns arising from unpurified PCR products were explained. It turns out, that the nondenaturing aqueous media assure both efficient DNA intake, and single-strand conformers stability required for the SSCP and combined SSCP/HA. The results of this study are applicable to all these areas of biomedical research, in which capillary electrophoresis is used for the characterization of nucleic acids.

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DNAElectrophoresis, CapillaryPolymerase Chain ReactionPolymorphism, Single NucleotidePolymorphism, Single-Stranded ConformationalJournal ArticleResearch Support, Non-U.S. Gov't

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Piotr Kozlowski, Wlodzimierz J Krzyzosiak. 2004. Optimum sample medium for single-nucleotide polymorphism and mutation detection by capillary electrophoresis.. https://doi.org/10.1002/elps.200305782

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