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SSCP/Heteroduplex Analysis

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PCR Mutation Detection Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 187))

Abstract

Single-strand conformation (SSCP) and heteroduplex analysis are separate mutation scanning methods in their own right. They are, however, unusual in that they can be carried out simultaneously on a single gel.

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References

  1. Orita, M., Iwahana, H., Kanazawa, H., Hayashi, K., and Sekiya, T. (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc. Natl. Acad. Sci. USA 86, 2766–2770.

    Article  PubMed  CAS  Google Scholar 

  2. Tassabehji, M., Newton, V. E., Turnbull, K., Seemanova, E., Kunze, J., Sperling, K., et al. (1994) PAX3 gene structure and mutations: close analogies between Waardenburg syndrome and the Splotch mouse. Hum. Mol. Genet. 7, 1069–1074.

    Article  Google Scholar 

  3. Makino, R., Yazyu, H., Kishimoto, Y., Sekiya, T., Hayashi, K. (1992) F-SSCP: fluorescence-based polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis. PCR Meth. Applic. 2, 10–13.

    CAS  Google Scholar 

  4. Hongyo, T., Buzard, G. S., Calvert, R. J., and Weghorst, C. M. (1993) ‘Cold SSCP’: a simple, rapid and non-radioactive method for optimized single-strand conformation polymorphism analyses. Nucl. Acids Res. 21, 3637–3642.

    Article  PubMed  CAS  Google Scholar 

  5. Sheffield, V. C., Beck, J. S., Kwitek, A. E., Sandstrom, D. W., and Stone, E. M. (1993) The sensitivity of single-strand conformation polymorphism analysis for the detection of single base substitutions. Genomics 16, 325–332.

    Article  PubMed  CAS  Google Scholar 

  6. White, M. B., Carvalho, M., Derse, D., O’Brien, S. J., and Dean, M. (1992) Detecting single base substitutions as heteroduplex polymorphisms. Genomics 12, 301–306.

    Article  PubMed  CAS  Google Scholar 

  7. Ganguly, A., Rock, M. J., and Prockop, D. J. (1993) Conformation-sensitive gel electrophoresis for rapid detection of single-base differences in double-stranded PCR products and DNA fragments: evidence for solvent-induced bends in DNA heteroduplexes. Proc. Natl. Acad. Sci. USA 90, 10,325–10,329.

    Article  PubMed  CAS  Google Scholar 

  8. Keen, J., Lester, D., Inglehearn, C, Curtis, A., and Bhattacharya, S. (1991) Rapid detection of single base mismatches as heteroduplexes on Hydrolink gels. Trends Genet. 7, 5.

    Article  PubMed  CAS  Google Scholar 

  9. Ravnik-Glavac, M., Glavac, D., and Dean, M. (1994) Sensitivity of single-strand conformation polymorphism and heteroduplex method for mutation detection in the cystic fibrosis gene. Hum. Mol. Genet. 3, 801–807.

    Article  PubMed  CAS  Google Scholar 

  10. Lee, H.-H., Lo, W.-J., and Choo, K.-B. (1992) Mutational analysis by a combined application of the multiple restriction fragment-single strand conformation polymorphism and the direct linear amplification sequencing protocols. Anal. Biochem. 205, 289–293.

    Article  PubMed  CAS  Google Scholar 

  11. Inazuka, M., Wenz, H. M., Sakabe, M., Tahira, T., and Hayashi, K. (1997) A streamlined mutation detection sysytem: multicolor post-PCR fluorescence labeling and single-strand conformational polymorphism analysis by capillary electrophoresis. Genome Res. 7, 1094–1103.

    PubMed  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. DNA Laboratory, Department of Medical Genetics, St. Mary’s Hospital, Manchester, UK

    Andrew J. Wallace

Authors
  1. Andrew J. Wallace
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Editor information

Editors and Affiliations

  1. Department of Haematology, Birmingham Children’s Hospital NHS Trust, Birmingham, UK

    Bimal D. M. Theophilus

  2. Department of Biosciences, University of Hertfordshire, Hatfield, UK

    Ralph Rapley

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© 2002 Humana Press Inc., Totowa, NJ

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Wallace, A.J. (2002). SSCP/Heteroduplex Analysis. In: Theophilus, B.D.M., Rapley, R. (eds) PCR Mutation Detection Protocols. Methods in Molecular Biology, vol 187. Humana Press. https://doi.org/10.1385/1-59259-273-2:151

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  • DOI: https://doi.org/10.1385/1-59259-273-2:151

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-617-8

  • Online ISBN: 978-1-59259-273-9

  • eBook Packages: Springer Protocols

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