Abstract
Detection of mutations is of central importance in the study of genetic and malignant diseases. Mutation detection helps us in understanding the protein structure, function, and expression. More than that, it is also important for presymptomatic/antenatal diagnosis, confirmation of the genetic cause of the disease and the mode of inheritance of a disease in a particular family, the prediction of clinical phenotype, and the potential for diagnostic analysis in the case of families with incomplete pedigrees or with new mutations. Therefore, the importance of direct mutation analysis cannot be understated.
Keywords
- Polymerase Chain Reaction Product
- BRCA1 Mutation
- Ethidium Bromide Solution
- Genetic Susceptibility Testing
- Polymerase Chain Reaction Optimization
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.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Hall, J. M., Friedman, L., Guenther, C, et al. (1990) Linkage of early-onset familial breast cancer to chromosome 17q21. Science 250, 1684–1689.
Miki, Y., Swensen, J., Shattuck-Eidens, D., et al. (1994) A strong candidate for breast and ovarian susceptibility gene BRCA1. Science 266, 66–71.
Wooster, R, Neuhausen, S. L., Mangoin, J., et al. (1994) Localization of a breast cancer susceptibility gene, BRCA2, to chromosome 13q12–13. Science 265, 2088–2090.
Wooster, R., Bignell, G, Lancaster, J., et al. (1995) Identification of breast cancer susceptibility gene BRCA2. Nature 378, 789–792.
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.
Cotton, R. G. H. (1993) Current methods of mutation detection. Mutat. Res. 285, 125–144.
Novack, D. F., Casna, N. J., Fischer, S. G., and Ford, J. P. (1986) Detection of single base-pair mismatches in DNA by chemical modification followed by electrophoresis in 15% polyacrylamide gel. Proc. Natl. Acad. Sci. USA 83, 586–590.
Myers, R. M., Larin, Z., and Maniatis, T. (1985) Detection of single base substitutions by ribonuclease cleavage at mismatches in RNA∶DNA duplexes. Science. 230, 1242–1246.
Cotton, R. G. H., Rodrigues, N. R., and Campbell, R. D. (1988) Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations. Proc. Natl. Acad. Sci. USA 85, 4397–4401.
Ganguly, A., Rooney, J. E., Hosomi, S., Zeiger, A., and Prockop, D. J. (1989) Detection and location of single-base mutations in large DNA fragments by immunomicroscopy. Genomics 4, 530–538.
Ganguly, A. and Prockop, D. J. (1990) Detection of single-base mutations by reaction of DNA heteroduplexes with a water-soluble carbodiimide followed by primer extension: application to products from the polymerase chain reaction. Nucleic Acids Res. 18, 3933–3939.
Youil, R., Kemper, B. W, and Cotton, R. G. (1995) Screening for mutations by enzyme mismatch cleavage with T4 endonuclease VII. Proc. Natl. Acad. Sci. USA 92, 87–91.
Myers, R. M., Maniatis, T., and Lerman, L. S. (1987) Detection and localization of single base changes by denaturing gradient gel electrophoresis Meth. Enzymol. 155, 501–527.
Sheffield, V. C., Cox, D. R., Lerman, L. S., and Myers, R. M. (1989) Attachment of a 40-base-pair G+C-rich sequence (GC-clamp) to genomic DNA fragments by the polymerase chain reaction results in improved detection of single-base changes. Proc. Natl. Acad. Sci. USA 86, 232–236.
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, 10325–10329.
Williams, C. J., Rock, M., Considene, E., McCarron, S., Gow, P., and Ladda, R. (1995) Three new point mutations in type II procollagen (COL2A1) and identification of a fourth family with the COL2A1 Arg519 to Cys base substitution using conformation sensitive gel electrophoresis. Hum. Mol. Genet. 4, 309–312.
Ganguly, T., Dhulipala, R., Godmilow, L., and Ganguly, A. (1998) High throughput fluorescence-based conformation-sensitive gel electrophoresis (F-CSGE) identifies six unique BRCA2 mutations and an overall low incidence of BRCA2 mutations in high-risk BRCA1-negative breast cancer families. Hum. Genet. 102, 549–556.
Korkoo, J., Pihlajamaa, T., Prockop, D. J., and Ala Kokko, L. (1996) Comparision of conformation sensitive gel electrophoresis with denaturing gel electrophoresis for detection of single base mutations in PCR products. Sixth International Conference on Matrix Biology, III-3.
Kumar, B. V., Lakhotia, S., Ankathil, R., Madhavan, J., Jayaprakash, P. G., Nair, K., and Somasundaram, K. (2002) Germline BRCA1 mutation analysis in Indian breast/ovarian cancer families. Cancer Biology and Therapy, in press.
Ganguly, A., Leahy, K., Marshall, A., Dhulipala, R., Godmilow, L., and Ganguly, T. (1997) Genetic testing for breast cancer susceptibility: frequency of BRCA1 and BRCA2 mutations. Genet. Testing 1, 85–90.
Schoettlin, W., Nielson, K. B., and Mathur, E. (1993) Strategies 6, 43–44.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Lakhotia, S., Somasundaram, K. (2003). Conformation-Sensitive Gel Electrophoresis for Detecting BRCA1 Mutations. In: El-Deiry, W.S. (eds) Tumor Suppressor Genes. Methods in Molecular Biology™, vol 223. Humana Press. https://doi.org/10.1385/1-59259-329-1:403
Download citation
DOI: https://doi.org/10.1385/1-59259-329-1:403
Publisher Name: Humana Press
Print ISBN: 978-0-89603-987-2
Online ISBN: 978-1-59259-329-3
eBook Packages: Springer Protocols