The SSCP method is based on the principle that the electrophoretic mobility of single-stranded DNA in a nondenaturing condition depends not only on its size but also on its sequence. Single-stranded DNA has a folded structure that is determined by intramolecular interactions.
This sequence-based secondary structure affects the mobility of the DNA during electrophoresis under nondenaturing conditions. A DNA molecule containing a mutation, even a single-base substitution, will have a different secondary structure than the wild type, resulting in a different mobility shift during electrophoresis than that of the wild type.
The technological aspects of CE-SSCP as well as the potential of CE-SSCP for routine genetic analysis were reviewed by Kourkine et al. in 2002.
Some examples of applications of SSCP by CGE include analysis of the p53 tumor suppressor gene for DNA diagnosis of cancer, detection of mutations on the K-RAS oncogene, detection of a Mycobacterium tuberculosis– specific amplified DNA fragment, detection of mutations in the β-globin gene promoter and the lipoprotein lipase and breast cancer 2 (BRCA2) genes, and identification of multiple mycobacterial species in a sample.
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