Comparative sensitivity of alternative single-strand conformation polymorphism (SSCP) methods.

摘要: Single-strand conformation polymorphism (SSCP) analysis is a genetic screening technique that allows for the rapid detection of single nucleotide substitutions in fragments PCR-amplified genomic DNA or cDNA. Several alternative protocols are now being commonly used to resolve differences electrophoretic mobility single-stranded fragments. The aim this study was directly compare sensitivity three popular SSCP methods using panel 19 known human mutations/polymorphisms present samples. Using electrophoresis protocol, 95% mutations were detected small-format PhastGel and PhastSystem. Large-format gels (5% polyacrylamide Hydrolink-MDE) tested both with without addition 10% glycerol. Hydrolink-MDE glycerol 89% 79%, respectively, 68% 63%, glycerol-containing gels. However, all either when non-glycerol examined. We conclude comparable very high efficiency can be achieved PhastSystem by combination two large-gel conditions Hydrolink-MDE.