A Nonradioactive Method for the Detection of Single-Strand Conformational Polymorphisms (SSCP)
The diagnosis of disease by direct analysis is an increasingly important branch of laboratory medicine. The detection of DNA sequence anomalies in different at-risk individuals may be effected by indirect techniques (e.g., linkage analysis), or by more direct methods involving the detection of sequence deviation. Large insertions or deletions can be detected by pulsed field gel electrophoresis or by using Southern blotting/RFLP techniques, however the more subtle alterations in DNA sequence such as point mutations or small deletions/insertions are below the level of resolution of these techniques. Several protocols have been reported that now allow detection of small deviations in DNA sequence (1, 2). Among the most promising of these is the technique for the detection of Single-Strand Conformational Polymorphisms (SSCPs).
KeywordsCystic Fibrosis Silver Nitrate Solution Subtle Alteration SSCP Variant MgCl2 Concentrate
- 5.Dockhorn-Dworniczak, B., Dworniczak, B., Brommellkamp, L., Bulles, J., Horst, J., and Bocker, W (1991) Non-isotopic detection of single-strand con-formational polymorphism (PCR-SSCP)-a rapid and sensitive technique in diagnosis of phenylketonuria Nucleic Acids Res. 19, 2500.PubMedCrossRefGoogle Scholar
- 11.Ainsworth, P.J., and Coulter-Mackie, M. B., (1990) A novel mutation in the gene for the a subunit of P-A′-acetylhexosaminidase associated with the B1 variant of Tay-Sachs disease. Amer. J. Hum. Genet. 47(Suppl.), 810.Google Scholar