Optimization of single-strand conformation polymorphism analysis in the presence of polyethylene glycol.
We report optimization of single-strand conformation polymorphism (SSCP) analysis in the presence of polyethylene glycol. The protocol developed separates single-strand conformers in a much shorter time (1-3 h) than conventional SSCP protocols and broadens the applicability of SSCP analysis from 150 to as much as 500 bp of DNA by different percentages of GC content present. We conclude that addition of polyethylene glycol helps improve the differential separation of conformers and, in combination with high-resolution polyacrylamide gel electrophoresis, offers an alternative to previous SSCP analysis protocols. This protocol should be very useful for clinical applications in routine detection of mutations as well as for research purposes.
Markoff
A
A
Laboratory of Molecular Biology and Genetics, Department of Chemistry and Biochemistry, Medical University Sofia, Bulgaria. markoff@uni-muenster.de
Savov
A
A
Vladimirov
V
V
Bogdanova
N
N
Kremensky
I
I
Ganev
V
V
D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005091:Exons; D019398:Genes, BRCA1 / Q000235:genetics; D006801:Humans; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011973:Receptors, LDL / Q000235:genetics
C505032:CFTR protein, human; D011973:Receptors, LDL; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D011092:Polyethylene Glycols; D004247:DNA