Skip to main content

Comparative Genomic Hybridization Technique

  • Protocol
Book cover Colorectal Cancer

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 50))

  • 710 Accesses

Abstract

Screening for chromosomal changes in solid tumors was long hindered by methodological problems encountered in standard cytogenetic analysis. Comparative genomic hybridization (CGH), a technique that emerged in 1992 (1) has proved to be a powerful tool for molecular cytogenetic analysis of neoplasms. The main prerequisite of the technique is DNA isolated from tumor samples. As no cell culture of tumor material is required, the technique has been successfully used to study fresh and frozen tissue samples, as well as archival formalin-fixed paraffin-embedded tissue samples. CGH allows to screen entire tumor genomes for gains and losses of DNA copy number, enabling consequent mapping of aberrations to chromosomal subregions. The technique is based on fluorescence in situ hybridization. Tumor and reference DNA are differentially labeled with fluorochromes (green and red, respectively) and mixed in equal amounts. The mixture is cohybridized competitively to a normal metaphase slide prepared from a lymphocyte cell culture of a normal healthy individual. After hybridization and washes, the chromosomes are counterstained with DAPI (blue) and slides are mounted with an antifading medium.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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

References

  1. Kallioniemi, A., Kallioniemi, O.-P., Sudan, D., Rutovitz, D., Gray, J., Waldman, F., and Pinkel, D. (1992) Comparative genomic hybridization for molecular cyto-genetic analysis of solid tumors. Science 258, 818–821.

    Article  CAS  PubMed  Google Scholar 

  2. Kallioniemi, O. P., Kallioniemi, A., Piper, J., Isola, J., Waldman, F. M., Gray, J. W., and Pinkel, D. (1994) Optimizing comparative genomic hybridization for analysis of DNA sequence copy number changes in solid tumors. Genes Chromosom. Cancer 10, 231–243.

    Article  CAS  PubMed  Google Scholar 

  3. Larramendy, M. L., El-Rifai, W., and Knuutila, S. (1998) Comparison of fluo-rescein isothiocyanate-and Texas red-conjugated nucleotides for direct labeling in comparative genomic hybridization. Cytometry 31, 174–179.

    Article  CAS  PubMed  Google Scholar 

  4. du Manoir, S., Schrock, E., Bentz, M., Speicher, M. R., Joos,., Ried, T., Lichter, P., and Cremer, T. (1995) Quantitative analysis of comparative genomic hybridization. Cytometry 19, 27–41.

    Article  PubMed  Google Scholar 

  5. El-Rifai, W., Larramendy, M. L., Björkqvist, A.-M., Hemmer, S., and Knuutila, S. (1997) Optimization of comparative genomic hybridization using 33 fluorochrome conjugated to dCTP and dUTP nucleotides. Lab. Invest. 77, 699–700.

    CAS  PubMed  Google Scholar 

  6. Knuutila, S., Björkqvist, A.-M., Autio, K., Tarkkanen, M., Wolf, M., Monni, O., et al. (1998) DNA copy number amplifications in human neoplasms. Review of comparative genomic hybridization studies. Am. J. Pathol. 152, 1107–1123.

    CAS  PubMed  Google Scholar 

  7. Knuutila, S., Aalto, Y., Autio, K., Björkqvist, A.-M., El-Rifai, W., Hemmer, S., et al. (1999) DNA copy number losses in human neoplasms. Am. J. Pathol. 155, 683–694.

    Article  CAS  PubMed  Google Scholar 

  8. Kokkola, A., Monni, O., Puolakkainen, P., Larramendy, M. L., Victorzon, M., Nordling, S., et al. (1997) 17q12-21 amplicon, a novel recurrent genetic change in intestinal type of gastric carcinoma: a comparative genomic hybridization study. Genes Chromosom. Cancer 20, 38–43.

    Article  CAS  PubMed  Google Scholar 

  9. El-Rifai, W., Harper, J. C., Cummings, O. W., Hyytinen, E. R., Frierson, H. F., Jr., Knuutila, S., and Powell, S. M. (1998) Consistent genetic alterations in xenografts of proximal stomach and gastro-esophageal junction adenocarcinomas. Cancer Res. 58, 34–37.

    CAS  PubMed  Google Scholar 

  10. van Dekken, H., Geelen, E., Dinjens, W. N., Wijnhoven, B. P., Tilanus, H. W., Tanke, H. J., and Rosenberg, C. (1999) Comparative genomic hybridization of cancer of the gastroesophageal junction: deletion of 14q31-32.1 discriminates between esophageal (Barrett′s) and gastric cardia adenocarcinomas. Cancer Res. 59, 748–52.

    PubMed  Google Scholar 

  11. Ried, T., Knutzen, R., Steinbeck, R., Blegen, H., Schrock, E., Heselmeyer, K., et al. (1996) Comparative genomic hybridization reveals a specific pattern of chromosomal gains and losses during the genesis of colorectal tumors. Genes Chromosom. Cancer 15, 234–245.

    Article  CAS  PubMed  Google Scholar 

  12. Al-Mulla, F., Keith, W. N., Pickford, I. R., Going, J. J., and Birnie, G. D. (1999) Comparative genomic hybridization analysis of primary colorectal carcinomas and their synchronous metastases. Genes Chromosom. Cancer 24, 306–314.

    Article  CAS  PubMed  Google Scholar 

  13. Sakakura, C., Mori, T., Sakabe, T., Ariyama, Y., Shinomiya, T., Date, K., et al. (1999) Gains, losses, and amplifications of genomic materials in primary gastric cancers analyzed by comparative genomic hybridization. Genes Chromosom. Cancer 24, 299–305.

    Article  CAS  PubMed  Google Scholar 

  14. Larramendy, M. L., El-Rifai, W., Kokkola, A., Puolakkainen, P., Monni, O., Salovaara, R., et al. (1998) Comparative genomic hybridization reveals differences in DNA copy number changes between sporadic gastric carcinoma and gastric carcinomas from patients with hereditary nonpolyposis colorectal cancer. Cancer Genet. Cytogenet. 106, 62–65.

    Article  CAS  PubMed  Google Scholar 

  15. El-Rifai, W., Sarlomo-Rikala, M., Knuutila, S., and Miettinen, M. (1998) DNA copy number changes in development and progression in leiomyosarcoma of soft tissues. Am. J. Pathol. 153, 985–990.

    Article  CAS  PubMed  Google Scholar 

  16. Hemminki, A., Tomlinson, I., Markie, D., Jarvinen, H., Sistonen, P., Bjorkqvist, A. M., et al. (1997) Localization of a susceptibility locus for Peutz-Jeghers syndrome to 19p using comparative genomic hybridization and targeted linkage analysis. Nature Genet. 15, 87–90.

    Article  CAS  PubMed  Google Scholar 

  17. Hemminki, A., Markie, D., Tomlinson, I., Avizienyte, E., Roth, S., Loukola, A., et al. (1998) A serine/threonine kinase gene defective in Peutz-Jeghers syndrome. Nature 391, 184–187.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2001 Humana Press Inc.

About this protocol

Cite this protocol

El-Rifai, W., Knuutila, S. (2001). Comparative Genomic Hybridization Technique. In: M., S. (eds) Colorectal Cancer. Methods in Molecular Medicine, vol 50. Humana Press. https://doi.org/10.1385/1-59259-084-5:25

Download citation

  • DOI: https://doi.org/10.1385/1-59259-084-5:25

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-767-0

  • Online ISBN: 978-1-59259-084-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics