Brief Communication | Published:

Analysis of repetitive DNA in chromosomes by flow cytometry

Nature Methods volume 8, pages 484486 (2011) | Download Citation

Abstract

We developed a flow cytometry method, chromosome flow fluorescence in situ hybridization (FISH), called CFF, to analyze repetitive DNA in chromosomes using FISH with directly labeled peptide nucleic acid (PNA) probes. We used CFF to measure the abundance of interstitial telomeric sequences in Chinese hamster chromosomes and major satellite sequences in mouse chromosomes. Using CFF we also identified parental homologs of human chromosome 18 with different amounts of repetitive DNA.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    et al. Blood 83, 1922–1928 (1994).

  2. 2.

    , , , & Methods 9, 113–121 (1996).

  3. 3.

    , , & Hum. Genet. 100, 138–144 (1997).

  4. 4.

    & Cytometry 11, 208–218 (1990).

  5. 5.

    , , , & Nature 293, 374–376 (1981).

  6. 6.

    et al. Nucleic Acids Res. 30, 2790–2799 (2002).

  7. 7.

    et al. Hum. Genet. 76, 290–292 (1987).

  8. 8.

    et al. Hum. Mol. Genet. 5, 685–691 (1996).

  9. 9.

    , , , & Nat. Biotechnol. 16, 743–747 (1998).

  10. 10.

    , , & Nat. Protoc. 1, 2365–2376 (2006).

  11. 11.

    & J. Histochem. Cytochem. 29, 74–78 (1981).

  12. 12.

    et al. Chromosoma 99, 3–10 (1990).

  13. 13.

    et al. Nature 463, 93–97 (2010).

  14. 14.

    , , & Nucleic Acids Res. 14, 2059–2073 (1986).

  15. 15.

    , , , & Nucleic Acids Res. 19, 1179–1182 (1991).

  16. 16.

    et al. Cell 117, 873–886 (2004).

  17. 17.

    , , , & Proc. Natl. Acad. Sci. USA 76, 1382–1384 (1979).

  18. 18.

    in Flow Cytogenetics (ed., J.W. Gray) 43–60 (Academic Press Inc., 1989).

  19. 19.

    , , & Cytometry 36, 267–278 (1999).

Download references

Acknowledgements

We thank B. Harrison, G. de Jong, L. Marmolejo and W. Xu for help with flow cytometry, and G. de Jong, S. Vanderbyl and L. Chavez for help with preparation of chromosome suspensions. HT1080 human fibrosarcoma cells (clone B49.5) were provided by W. Bickmore (Medical Research Council Human Genetics Unit). Work in the Lansdorp laboratory is supported by grants from the Canadian Institutes of Health Research (RMF-92093), the US National Institutes of Health (R01GM094146), the Canadian Cancer Society and the Terry Fox Foundation (018006 and 105265).

Author information

Affiliations

  1. Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia, Canada.

    • Julie Brind'Amour
    •  & Peter M Lansdorp
  2. Division of Hematology, Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada.

    • Peter M Lansdorp
  3. European Research Institute on the Biology of Ageing, University of Groningen, University Medical Centre, Groningen, The Netherlands.

    • Peter M Lansdorp

Authors

  1. Search for Julie Brind'Amour in:

  2. Search for Peter M Lansdorp in:

Contributions

J.B. performed experiments designed by J.B. and P.M.L.; and J.B. and P.M.L. wrote the paper.

Competing interests

P.M.L. is a founding shareholder in Repeat Diagnostics Inc., a company specializing in leukocyte telomere length measurements using flow FISH.

Corresponding author

Correspondence to Peter M Lansdorp.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–8 and Supplementary Note 1

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/nmeth.1601

Further reading