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Analysis of repetitive DNA in chromosomes by flow cytometry

Nature Methods volume 8, pages 484–486 (2011)Cite this article

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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.

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Figure 1: Preservation of flow karyograms after FISH enables chromosome-specific detection of interstitial telomere repeats in CHO cell chromosomes.
Figure 2: Chromosome and allele-specific analysis of satellite DNA in mouse and human cell lines.

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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).

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Authors and 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

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  1. Julie Brind'Amour
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  2. Peter M Lansdorp
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Contributions

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

Corresponding author

Correspondence to Peter M Lansdorp.

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Competing interests

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

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Supplementary Figures 1–8 and Supplementary Note 1 (PDF 2053 kb)

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Brind'Amour, J., Lansdorp, P. Analysis of repetitive DNA in chromosomes by flow cytometry. Nat Methods 8, 484–486 (2011). https://doi.org/10.1038/nmeth.1601

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