Advertisement

Modifications to SSCP Analysis Conditions

  • William C. Jordan
  • Katherine Foley
  • Michael W. Bruford
Chapter

Abstract

The electrophoretic conditions under which SSCPs can be detected must be determined empirically for each set of mutations under analysis. However, a number of parameters has been identified as important in determining the sensitivity of SSCP detection, as follows.

Keywords

Buffer Concentration Screening Protocol SSCP Analysis Transverse Temperature Mobility Difference 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Orita, M., H. Iwahana, H. Kanazawa, K. Hayashi, and T. Sekiya (1989a) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proceedings of National Academy of Sciences. USA 86, 2766–2770.CrossRefGoogle Scholar
  2. 2.
    Orita, M., Y. Suzuki, T. Sekiya, and K. Hayashi (1989b) Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5, 874–879.CrossRefGoogle Scholar
  3. 3.
    Lessa, E. P. and G. Applebaum (1993) Screening techniques for detecting allelic variation in DNA sequences. Molecular Ecology 2, 119–129.CrossRefGoogle Scholar
  4. 4.
    Ravnik-Glavač, M., D. Glavač, and M. Dean (1994) Sensitivity of single-strand conformation polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene. Human Molecular Genetics 3, 801–807.CrossRefGoogle Scholar
  5. 5.
    Hayashi, K. (1991) PCR-SSCP: a simple and sensitive method for detection of mutations in the genomic DNA. PCR Methods and Applications 1, 34–38.Google Scholar
  6. 6.
    Sheffield, V.C., J.S. Beck, A.E. Kwitek, and E.M. Stone (1993) The sensitivity of single-strand conformation polymorphism analysis for the detection of single base substitutions. Genomics 16, 325–332.CrossRefGoogle Scholar
  7. 7.
    Fan, E., D.B. Levin, B.W. Glickman, and D.M. Logan (1993) Limitations in the use of SSCP analysis. Mutation Research 288, 85–92.CrossRefGoogle Scholar
  8. 8.
    Liu, Q., and S.S. Sommer (1994) Parameters affecting the sensitivities of dideoxy finger-printing and SSCP. PCR Methods and Applications 4, 97–108.Google Scholar
  9. 9.
    Nielson, D.A., A. Novoradovsky, and D. Goldman (1995) SSCP primer design based on single-strand DNA structure predicted by a DNA folding program. Nucleic Acids Research. 23, 2287–2291.CrossRefGoogle Scholar
  10. 10.
    Iwahana, H., K. Yoshimoto, N Mizusawa, E. Kudo, and M Itakura (1994) Multiple fluorescence-based PCR-SSCP analysis. Bio-Techniques 16, 296–305.Google Scholar
  11. 11.
    Yap, E.P.H. and J.O’D. McGee (1992) Nonisotopic SSCP detection in PCR products by ethidium bromide staining. Trends in Genetics 8, 49.CrossRefGoogle Scholar
  12. 12.
    Chaubert, P., D. Bautista, and J. Benhattar (1993) An improved method for rapid screening of DNA mutations by non-radioactive single-strand conformation polymorphism procedure. Bio Techniques 15, 586.Google Scholar
  13. 13.
    Weghorst, C.M., and G.S. Buzard (1993) Enhanced single-strand conformation polymorphism (SSCP) detection of point mutations utilizing methylmercury hydroxide. Bio Techniques 15, 396–400.Google Scholar
  14. 14.
    Maruya, E., H. Saji, and S. Yokoyama (1996) PCR-LIS-SSCP (low ionic strength single-stranded conformation polymorphism) — a simple method for high resolution allele typing of HLA-DRB1, -DQB1, and -DPB1. Genome Research 6, 51–57.CrossRefGoogle Scholar
  15. 15.
    Russell, G.C. (1994) Improved single-strand conformation polymorphism analysis by asymmetric polymerase chain reaction with end-labelled primers. Genetic Analysis Techniques Applications 11, 24–27.CrossRefGoogle Scholar
  16. 16.
    Kasuga, T., J. Cheng, and K.R. Mitchelson (1995) Metastable single-strand DNA conformational polymorphism analysis results in enhanced polymorphism detection. PCR Methods and Applications 4, 227–233.Google Scholar
  17. 17.
    Monckton, D.G., and A.J. Jeffreys (1994) Minisatellite isoalleles can be distinguished by single-strand conformation polymorphism analysis in agarose gels. Nucleic Acids Research. 22, 2155–2157.CrossRefGoogle Scholar
  18. 18.
    Peng, H., M. Du, J. Ji, P.G. Isaacson, and L. Pan (1995) High-resolution SSCP analysis using Polyacrylamide agarose composite gel and a background-free silver staining method. Bio-Techniques 19, 410–414.Google Scholar
  19. 19.
    Spinardi, L., R. Mazars, and C. Theillet. (1991) Protocols for an improved detection of point mutations by SSCP. Nucleic Acids Research. 19, 4009.CrossRefGoogle Scholar
  20. 20.
    Chen, X., T. Baumstark, G. Steger, and D. Riesner (1995) High resolution SSCP by optimization of the temperature by transverse TGGE. Nucleic Acids Research. 23, 4524–4525.CrossRefGoogle Scholar
  21. 21.
    Grace, M.B., G.S. Bambeck, G.S Buzard, and B.D. Weintraub (1995) Tranverse temperature-gradient single-strand conformation polymorphism analysis for temperature optimization of ‘Cold’-SSCP mutation detection. Nucleic Acids Research. 23, 4224–4226.CrossRefGoogle Scholar

Copyright information

© Chapman & Hall 1998

Authors and Affiliations

  • William C. Jordan
  • Katherine Foley
  • Michael W. Bruford

There are no affiliations available

Personalised recommendations