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Clinical Chemistry 43: 30-33, 1997;
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(Clinical Chemistry. 1997;43:30-33.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Optimization of single-strand conformation polymorphism analysis in the presence of polyethylene glycol

Arseni Markoff1,2, Alex Savov2, Vladimir Vladimirov1, Nadia Bogdanova2, Ivo Kremensky2 and Varban Ganev1

1 Laboratory of Molecular Biology and Genetics, Department of Chemistry and Biochemistry, Medical University Sofia, 2 Zdrave Str., 1431 Sofia, Bulgaria.

2 Laboratory of Molecular Pathology, University Hospital of Obstetrics and Gynaecology, 2 Zdrave Str., 1431 Sofia, Bulgaria.

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.


Key Words: indexing terms: polymerase chain reaction • detection of mutations and polymorphisms • electrophoresis, polyacrylamide gel




The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
L. K. Hennessy, J. Teare, and C. Ko
PCR Conditions and DNA Denaturants Affect Reproducibility of Single-Strand Conformation Polymorphism Patterns for BRCA1 Mutations
Clin. Chem., April 1, 1998; 44(4): 879 - 882.
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Copyright © 1997 by the American Association for Clinical Chemistry.