Abstract
As an increasing number of human diseases are linked to the effects of altered genes, new methods are being sought for detection of mutations and their relationship to the presence of disease. Since total genomic DNA usually cannot be analyzed directly, target sequences are amplified by the polymerase chain reaction (PCR). Several methods have been reported that allow detection of small changes in DNA sequence (1). Among them, one of the most commonly used is the heteroduplex analysis method (HA) (2–5).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Cotton, R. G. H. (1993) Current methods of mutation detection. Mutat. Res. 285, 125–144.
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.
Perry, D. J. and Carrell, R. W. (1992) HydroLink gels: A rapid and simple approach to the detection of DNA mutations in thromboembolic disease. J. Clin. Pathol. 45, 158–160.
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.
Glavac, D. and Dean, M. (1995) Applications of heteroduplex analysis for mutation detection in disease genes. Hum. Mutat. 6, 281–287.
Mashal, R. D. and Sklar, J. (1996) Practical methods of mutation detection. Curr. Opin. Genet. Develop. 6, 275–280.
Cenarro, A., Jensen, H. K., Casao, E., Civeira, F., González-Bonillo, J., Pocoví, M., and Gregersen, N. (1996) Identification of a novel mutation in exon 13 of the LDL receptor gene causing familial hypercholesterolemia in two Spanish families. Biochim. Biophys. Acta 1316, 1–4.
Soto, D. and Sukumar, S. (1992) Improved detection of mutations in the p53 gene in human tumors as single-stranded conformation polymorphisms and double-stranded heteroduplex DNA. PCR Meth. Appl. 2, 96–98.
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.
Bhattacharya, A. and Lilley, D. M. (1989) The contrasting structures of mismatched DNA sequences containing looped-out bases (bulges) and multiple mismatches (bubbles). Nucleic Acids Res. 17, 6821–6840.
Williams, C. J., Rock, M., Considine, E., McCarron, S., Gow, P., Ladda, R., et al. (1995) Three new point mutations in type II procollagen (COL2A1) and identification of a fourth family with the COL2A1 Arg519→Cys base substitution using conformation sensitive gel electrophoresis. Hum. Mol. Genet. 4, 309–312.
Körkko, J., Annunen, S., Puilajamaa, T., Prockop, D. J., and Ala-Kokko, L. (1998) Conformation sensitive gel electrophoresis for simple and accurate detection of mutations: Comparison with denaturing gradient gel electrophoresis and nucleotide sequencing. Proc. Natl. Acad. Sci. USA 95, 1681–1685.
Williams, I. J., Abuzenadah, A., Winship, P. R., Preston, F. E., Dolan, G., Wright, J., et al. (1998) Precise carrier diagnosis in families with haemophilia A: use of conformation sensitive gel electrophoresis for mutation screening and polymorphism analysis. Thromb. Haemost. 79, 723–726.
Recalde, D., Cenarro, A., Civeira, F., and Pocoví, M. (1998) Apo A-I Zaragoza (L144R): A novel mutation in the apolipoprotein A-I gene associated with familial hypoalphalipoproteinemia. Hum. Mutat. Mutation and Polymorphism Report 11, 416.
Makino, R., Yazyu, H., Kishimoto, Y., Sekiya, T., and Hayashi, K. (1992) F-SSCP: Fluorescence-based polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis. PCR Meth. Appl. 2, 10–13.
Nagamine, C. M., Chan, K., and Lau, Y-F. C. (1989) A PCR artifact: Generation of heteroduplexes. Am. J. Hum. Genet. 45, 337–339.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Cenarro, A., Civeira, F., Pocovi, M. (1999). Analysis of Genetic Variants in Cardiovascular Risk Genes by Heteroduplex Analysis. In: Baker, A.H. (eds) Vascular Disease. Methods in Molecular Medicine™, vol 30. Humana Press. https://doi.org/10.1385/1-59259-247-3:13
Download citation
DOI: https://doi.org/10.1385/1-59259-247-3:13
Publisher Name: Humana Press
Print ISBN: 978-0-89603-731-1
Online ISBN: 978-1-59259-247-0
eBook Packages: Springer Protocols