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Characterization of Chromosomal Rearrangements Using Multicolor-Banding (MCB/m-band)

  • Thomas LiehrEmail author
  • Anja Weise
  • Sophie Hinreiner
  • Hasmik Mkrtchyan
  • Kristin Mrasek
  • Nadezda Kosyakova
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 659)

Abstract

Molecular cytogenetics and especially fluorescence in situ hybridization (FISH) banding approaches are nowadays standard for the exact characterization of simple, complex, and cryptic chromosomal aberrations within the human genome. FISH-banding techniques are any kind of FISH techniques, which provide the possibility to characterize simultaneously several chromosomal subregions smaller than a chromosome arm. FISH banding methods fitting that definition may have quite different characteristics, but share the ability to produce a DNA-specific chromosomal banding. While the standard techniques such as G-bands by Trypsin using Giemsa banding lead to a protein-related black and white banding pattern, FISH-banding techniques are DNA-specific, more colorful, and thus, more informative. At present, the most frequently applied FISH banding technique is the multicolor banding (MCB/m-band) approach. MCB/m-band is based on region-specific microdissection libraries, producing changing fluorescence intensity ratios along the chromosomes. Here we describe the FISH-banding technique MCB/m-band and illustrate how to apply it for characterization of chromosomal breakpoints with a minimal number of FISH experiments.

Key words

FISH-banding Multicolor banding Microdissection Marker chromosomes Chromosomal breakpoints 

Notes

Acknowledgments

Supported in parts by the DFG (436 RUS 17/109/04, 436 RUS 17/22/06, WE 3617/2-1, 436 ARM 17/5/06, LI820/11-1, LI820/17-1), Boehringer Ingelheim Fonds, Evangelische Studienwerk e.V. Villigst, IZKF Jena (Start-up S16), IZKF together with the TMWFK (TP 3.7 and B307-04004), Stiftung Leukämie, and Stefan-Morsch-Stiftung.

References

  1. 1.
    Seabright, M. (1971) A rapid banding technique for human chromosomes. Lancet 30, 971–972.CrossRefGoogle Scholar
  2. 2.
    Liehr, T., Weise, A., Heller, A., Starke, H., Mrasek, K., Kuechler, A., Weier, H. U., and Claussen, U. (2002) Multicolor chromosome banding (MCB) with YAC/BAC-based probes and region-specific microdissection DNA libraries. Cytogenet Genome Res 97, 43–50.PubMedCrossRefGoogle Scholar
  3. 3.
    Pinkel, D., Straume, T., and Gray, J. W. (1986) Cytogenetic analysis using quantitative, high-sensitivity, fluorescence hybridization. Proc Natl Acad Sci U S A 83, 2934–2938.PubMedCrossRefGoogle Scholar
  4. 4.
    Chang, S. S., and Mark, H. F. L. (1997) Emerging molecular cytogenetic techniques. Cytobios 90, 7–22.PubMedGoogle Scholar
  5. 5.
    Speicher, M. R., Gwyn Ballard, S., and Ward, D. C. (1996) Karyotyping human chromosomes by combinatorial multi-fluor FISH. Nat Genet 12, 368–375.PubMedCrossRefGoogle Scholar
  6. 6.
    Schröck, E., du Manoir, S., Veldman, T., Schoell, B., Wienberg, J., Ferguson-Smith, M. A., Ning, Y., Ledbetter, D. H., Bar-Am, I., Soenksen, D., Garini, Y., and Ried, T. (1996) Multicolor spectral karyotyping of human chromosomes. Science 273, 494–497.PubMedCrossRefGoogle Scholar
  7. 7.
    Liehr, T., Starke, H., Heller, A., Kosyakova, N., Mrasek, K., Gross, M., Karst, C., Steinhaeuser, U., Hunstig, F., Fickelscher, I., Kuechler, A., Trifonov, V., Romanenko, S. A., and Weise, A. (2006) Multicolor fluorescence in situ hybridization (FISH) applied to FISH-banding. Cytogenet Genome Res 114, 240–244.PubMedCrossRefGoogle Scholar
  8. 8.
    Liehr, T., Gross, M., Karst, C., Glaser, M., Mrasek, K., Starke, H., Weise, A., Mkrtchyan, H., and Kuechler, A. (2006) FISH banding in tumor cytogenetics. Cancer Genet Cytogenet 164, 88–89.PubMedCrossRefGoogle Scholar
  9. 9.
    Weise, A., Mrasek, K., Fickelscher, I., Claussen, U,. Cheung, S. W., Cai, W. W., Liehr, T., and Kosyakova, N. (2008) Molecular definition of high-resolution multicolor banding probes: first within the human DNA sequence anchored FISH banding probe set. J Histochem Cytochem 56, 487–493.PubMedCrossRefGoogle Scholar
  10. 10.
    Chudoba, I., Plesch, A., Lörch, T., Lemke, J., Claussen, U., and Senger, G. (1999) High resolution multicolor-banding: a new technique for refined FISH analysis of human chromosomes. Cytogenet Cell Genet 84, 156–160.PubMedCrossRefGoogle Scholar
  11. 11.
    Liehr, T., Heller, A., Starke, H., Rubtsov, N., Trifonov, V., Mrasek, K., Weise, A., Kuechler, A., and Claussen, U. (2002) Microdissection based high resolution multicolor banding for all 24 human chromosomes. Int J Mol Med 9, 335–339.PubMedGoogle Scholar
  12. 12.
    Weise, A., Heller, A., Starke, H., Mrasek, K., Kuechler, A., Pool-Zobel, B. L., Claussen, U., and Liehr, T. (2003) Multitude multicolor chromosome banding (mMCB) – a comprehensive one-step multicolor FISH banding method. Cytogenet Genome Res 103, 34–39.PubMedCrossRefGoogle Scholar
  13. 13.
    Lemke, J., Claussen, J., Michel, S., Chudoba, I., Mühlig, P., Westermann, M., Sperling, K., Rubtsov, N., Grummt, U.W., Ullmann, P., Kromeyer-Hauschild, K., Liehr, T., and Claussen, U. (2002) The DNA-based structure of human chromosome 5 in interphase. Am J Hum Genet 71, 1051–1059.PubMedCrossRefGoogle Scholar
  14. 14.
    Iourov, I. Y., Liehr, T., Vorsanova, S. G., Kolotii, A. D., and Yurov, Y. B. (2006) Visualization of interphase chromosomes in postmitotic cells of the human brain by multicolour banding (MCB). Chromosome Res 14, 223–229.PubMedCrossRefGoogle Scholar
  15. 15.
    Liehr, T., and Claussen, U. (2002) FISH on chromosome preparations of peripheral blood, in: FISH-Technology (Rautenstrauss, B. and Liehr, T., eds.) Springer, Berlin, pp 73–81.CrossRefGoogle Scholar
  16. 16.
    Liehr, T., Thoma, K., Kammler, K., Gehring, C., Ekici, A., Bathke, K. D., Grehl, H., and Rautenstrauss, B. (1995) Direct preparation of uncultured EDTA-treated or heparinized blood for interphase FISH analysis. Appl Cytogenet 21, 185–188Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Thomas Liehr
    • 1
    Email author
  • Anja Weise
    • 1
  • Sophie Hinreiner
    • 1
  • Hasmik Mkrtchyan
    • 2
  • Kristin Mrasek
    • 1
  • Nadezda Kosyakova
    • 1
  1. 1.Institut für Humangenetik und AnthropologieUniversity of JenaJenaGermany
  2. 2.Institut für HumangenetikUniversity of JenaJenaGermany

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