Skip to main content

Constant Denaturant Gel Electrophoresis (CDGE) in Mutation Screening

  • Chapter
Technologies for Detection of DNA Damage and Mutations

Abstract

There is an increasing need for practical, efficient, and inexpensive ways to explore inherited mutations responsible for genetic diseases, acquired mutations involved in cancer development, and induced mutations in mutational spectrometry. Most of the current methods used for mutation screening are based on PCR, and this has solved the problem of target limitation. Simple and efficient methods such as allele-specific hybridization, allele-specific amplification, ligation, primer extension and artificial introduction of restriction sites, and variants of these methods have been developed for screening large numbers of samples for one particular mutation (for review see Cotton, 1993).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Andersen, T. L., Holm, R., Nesland, J. M., Heimdal, K. R., Ottestad, L., and Børresen, A.-L. (1993). Prognostic significance of TP53 alterations in breast carcinoma. Br. J. Cancer 68:540–548.

    Article  PubMed  CAS  Google Scholar 

  • Andersson, B., Ying, J. H., Lewis, D. E., and Gibbs, R. A. (1993). Rapid characterization of HIV-1 sequence diversity using denaturing gradient gel electrophoresis and direct automated DNA sequence of PCR products. PCR Methods Appl. 2(4):293–300.

    Article  PubMed  CAS  Google Scholar 

  • Børresen, A.-L., Hovig, E., Smith-S—rensen, B., Malkin, D., Lystad, S., Andersen, T. I., Nesland, J. M., Isselbacher, K. J., and Friend, S. H. (1991). Constant dĂ©naturant gel electrophoresis as a rapid screening technique for p53 mutations. Proc. Natl. Acad. Sci. USA 88:8405–8409.

    Article  PubMed  Google Scholar 

  • Børresen, A.-L., Andersen, T. I., Garber, J., Piraux, N. B., Thorlacius, S., Eyfjrd, J., Ottestad, L., Smith-Sørensen, B., Hovig, E., Malkin, D., and Friend, S. H. (1992). Screening for germ line TP53 mutations in breast cancer patients. Cancer Res. 52:3234–3236.

    PubMed  Google Scholar 

  • Condie, A., Eeeles, R., Børresen, A.-L., Coles, C., Cooper, C., and Prosser, J. (1993). Detection of point mutations in the p53 gene. Comparison of single-strand conformation polymorphism, constant denaturant gel electrophoresis, and hydroxylamine and osmium tetroxide techniques. Hum. Mutat. 2:58–66.

    Article  PubMed  CAS  Google Scholar 

  • Cornelis, R. S., van Vliet, M., Vos, C. B. J., Cleton-Jansen, A.-M., van de Vrjver, M. J., Peterse, J. L., Meera Khan, P., Børresen, A.-L., Cornelisse, C. J., and Devilee, P (1994). Evidence for a gene on 17p 13.3, distal to TP53, as a target for allele loss in breast tumors without p53 mutations. Cancer 54:4200–4206.

    CAS  Google Scholar 

  • Costes, B., Girodon, E., Ghanem, N., Chassignol, M., Thuong, N.T., Dupret, D., and Goossens, M. (1993). Psoralen-modified oligonucleotide primers improve detection of mutations by denaturing gradient gel electrophoresis and provide an alternative to GC-clamping. Hum. Mol. Genet. 2(4):393–397.

    Article  PubMed  CAS  Google Scholar 

  • Cotton, R. G. H. (1993). Current methods of mutation detection. Mutat. Res. 285:125–144.

    Article  PubMed  CAS  Google Scholar 

  • Dworniczak, B., Kalydjieva, L., Pankoke, S., Aulehla-Scholz, C., Allen, G., and Horst, J. (1992). Analysis of exon 7 of the human phenylalanine hydroxylase gene: A mutation hot spot? Hum. Mutat. 1(2):138–146.

    Article  PubMed  CAS  Google Scholar 

  • Fischer, S. G., and Lerman, L. S. (1983). DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: Correspondence with melting theory. Proc. Natl. Acad. Sci. USA 80:1579–1583.

    Article  PubMed  CAS  Google Scholar 

  • Gotoh, O., and Tagashira, Y. (1981). Location of frequently opening regions of natural DNA’s and their relation to functional loci. Biopolymers 20(5):1043–1058.

    Article  PubMed  CAS  Google Scholar 

  • Guldberg, P., Henriksen, K. F., and Guttler, F. (1993). Molecular analysis of phenylketonuria in Denmark:99% of the mutations detected by denaturing gradient gel electrophoresis. Genomics 17(1):141–146.

    Article  PubMed  CAS  Google Scholar 

  • Heimdal, K. R., Lothe, R. A., Lystad, S., Holm, R., Foss, S. D., and Børresen, A.-L. (1993). No germline TP53 mutations detected in familial and bilateral testicular cancer. Genes Chromosomes Cancer 6(2):92–97.

    Article  PubMed  CAS  Google Scholar 

  • Hovig, E., Smith-Sørensen, B., Brøgger, A., and Børresen, A.-L. (1991). Constant dĂ©naturant gel electrophoresis, a modification of denaturing gel electrophoresis, in mutation detection. Mutat. Res. 262:63–71.

    Article  PubMed  CAS  Google Scholar 

  • Hovig, E., Smith-Sørensen, B., Gebhardt, M. C., Ryberg, D., Lothe, R., and Børresen, A.-L. (1992). Alterations in retinoblastoma susceptibility gene exon 21 are not common in human tumours. Genes Chromosomes Cancer 5 (2):97–103.

    Article  PubMed  CAS  Google Scholar 

  • Hovig, E., Smith-Sørensen, B., and Børresen, A.-L. (1994). Detection of mutation by denaturing gradient gel electrophoresis, in:Current Protocols in Human Genetics (N. Dracopoli, D. Moir, D. Smith, J. Haines, C. Morton, B. Korf, C. Seideman, and J. Seidman, eds.) Current Protocols, New York, Chapter 7.5.

    Google Scholar 

  • Husgafvel-Pursiainen, K., Ridanpää, M., Hackman, P., Antilla, S., Karjalainen, A., Ă–nfelt, A., Børresen, A.-L., and Vainio, H. (1992). Detection of ras gene mutations in human lung cancer: Comparison of two screening assays based on the polymerase chain reaction. Environ. Health Perspect. 98:183–185.

    Article  PubMed  CAS  Google Scholar 

  • Khrapko, K., Hanekamp, J. S., Thilly, W. G., Belenki, A., Foret, F., and Karger, B. L. (1994). Constant denaturant capillary electrophoresis (CDCE):A high resolution approach to mutational analysis. Nucleic Acids Res. 22:364–372.

    Article  PubMed  CAS  Google Scholar 

  • Lerman, L. S., and Silverstein, K. (1987). Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods Enzymol. 155:482–501.

    Article  PubMed  CAS  Google Scholar 

  • Mclntyre, J. R., Smith-Sørensen, B., Friend, S. H., Kassell, J., Børresen, A.-L., Yu Xin Yan, Russo, C., Sato, J., Barbier, N., Miser, J., Malkin, D., and Gebhardt, M. C. (1994). Germline mutations of the p53 tumor suppressor gene in children with osteosarcoma. J. Clin. Oncol. 12:925–930.

    Google Scholar 

  • Malkin, D., Jolly, K. W., Barbier-Piraux, N., Look, T., Friend, S. H., Gebbhardt, M. C., Andersen, T. I., Børresen, A.-L., Li, F. P., Garber, J., and Strong, L. C. (1992). Germline mutations in the p53 tumor suppressor gene in children and young adults with second malignant neoplasms. N. Eng. J. Med. 20:1309–1315.

    Article  Google Scholar 

  • Peng, H. Q., Malkin, D., Bailey, D., Gallie, B. L., Bulbul, M., Jewet, M., Buchanan, J., and Goss, P. E. (1993). Mutations of the p53 genes do not occur in testis cancer. Cancer Res. 53:3574–3578.

    PubMed  CAS  Google Scholar 

  • Poland, D. (1974). Recursion relation generation of probability profiles for specific-sequence macromolecules with long-range correlations. Biopolymers 13:1859–1871.

    Article  PubMed  CAS  Google Scholar 

  • Ridanpää, M., and Husgafvel-Pursiainen, K. (1993). Denaturing gradient gel electrophoresis (DGGE) assay for K-ras and N-ras genes: Detection of K-ras point mutations in human lung tumour DNA. Hum. Mol. Genet. 2(6):639–644.

    Article  PubMed  Google Scholar 

  • Ryberg, D., Kure, E., Lystad, S., Skaug, V., Stangeland, L., Mercy, L, Børresen, A.-L., and Haugen, A. (1994). Mutations in lung tumors. Relationship to putative susceptibility markers for cancer. Cancer Res. 54:1551–1555.

    PubMed  CAS  Google Scholar 

  • Seruca, R., David, L., Holm, R., Nesland, J. M., Fangan, B. M., Castedo, S., Sobrinho-Simoes, M., and Børresen, A.-L. (1992). P53 mutations in gastric carcinomas. Br. J. Cancer 65(5):708–710.

    Article  PubMed  CAS  Google Scholar 

  • 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. Nad. Acad. Sci. USA 86:232–236.

    Article  CAS  Google Scholar 

  • Smith-Sørensen, B., Hovig, E., Andersson, B., and Børresen, A.-L. (1992). Screening for base mutations in human HPRT cDNA using the polymerase chain reaction (PCR) in combination with constant dĂ©naturant gel electrophoresis (CDGE). Mutat. Res. 269:41–53.

    Article  PubMed  Google Scholar 

  • Smith-Sørensen, B., Gebhardt, M. C., Kloen, P., Aguilar, F., Friend, S. H., and Børresen, A.-L. (1993). Screening for TP53 mutations in osteosarcomas using the polymerase chain reaction (PCR) in combination with constant denaturant gel electrophoresis (CDGE). Hum. Mutat. 2:274–285.

    Article  PubMed  Google Scholar 

  • Thorlacius, S., Børresen, A.-L., and Eyfjörd, J. (1993). Somatic p53 mutations in human breast carcinomas in an Icelandic population, a prognostic factor. Cancer Res. 53:1637–1641.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1996 Springer Science+Business Media New York

About this chapter

Cite this chapter

Børresen, AL. (1996). Constant Denaturant Gel Electrophoresis (CDGE) in Mutation Screening. In: Pfeifer, G.P. (eds) Technologies for Detection of DNA Damage and Mutations. Springer, Boston, MA. https://doi.org/10.1007/978-1-4899-0301-3_20

Download citation

  • DOI: https://doi.org/10.1007/978-1-4899-0301-3_20

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4899-0303-7

  • Online ISBN: 978-1-4899-0301-3

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics