Molecular Biotechnology

, Volume 27, Issue 2, pp 169–172 | Cite as

Real-time quantitative PCR of telomere length

  • Marcel E. GilEmail author
  • Thérèsa L. Coetzer
Hints and Tips


Telomeres cap the ends of chromosomes and are essential for the protection of chromosomes, as well as restricting the replicative potential of a cell. These functions are achieved by the regulation of telomeric repeat length, making the measurement of telomere length a useful aid in the elucidation of the replicative history and potential of cells. Previously published techniques employed either hybridization or flow cytometry methods, which are technically demanding and time-consuming. In 2002, R. M. Cawthon published a real-time polymerase chain reaction (PCR)-based method for telomere length measurement using the Applied Biosystems Prism 7700 sequence detection system. The technique measures the factor by which the ratio of telomere repeat copy number to single-gene copy number differs between a sample and that of a reference deoxyribonucleic acid sample. In many laboratories worldwide, including ours, real-time PCR is carried out using the Roche LightCycler, as opposed to the AB Prism 7700 system. This benchmark details the modifications to Cawthon’s method and describes the parameters and reagents required to measure telomere length using the Roche LightCycler.

Index Entries

LightCycler telomere length telomeres real-time PCR 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Cech, T. R. (2000) Life at the end of the chromosome: telomeres and telomerase. Angew. Chem. Int. Ed. Engl. 39, 34–43.PubMedCrossRefGoogle Scholar
  2. 2.
    Hahn, W. C. (2003) Role of telomeres and telomerase in the pathogenesis of human cancer. J. Clin. Oncol. 21, 2034–2043.PubMedCrossRefGoogle Scholar
  3. 3.
    Mathon, N. F. and Lloyd, A. C. (2001) Cell senescence and cancer. Nat. Rev. Cancer 1, 203–213.PubMedCrossRefGoogle Scholar
  4. 4.
    Harley, C. B., Futcher, A. B., and Greider, C. W. (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345, 458–460.PubMedCrossRefGoogle Scholar
  5. 5.
    Bryant, J. E., Hutchings, K. G., Moyzis, R. K., and Griffith, J. K. (1997) Measurement of telomeric DNA content in human tissues. Biotechniques 23, 476–484.PubMedGoogle Scholar
  6. 6.
    Nakamura, Y., Hirose, M., Matsuo, H., Tsuyama, N., Kamisango, K., and Ide, T. (1999) Simple, rapid, quantitative and sensitive detection of telomere repeats in cell lysate by a hybridisation protection assay. Clin. Chem. 45, 1718–1724.PubMedGoogle Scholar
  7. 7.
    Baerlocher, G. M., Mak, J., Tien, T., and Lansdorp, P. M. (2002) Telomere length measurement by fluorescence in situ hybridisation and flow cytometry: tips and pitfalls. Cytometry 47, 89–99.PubMedCrossRefGoogle Scholar
  8. 8.
    Cawthon, R. M. (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res. 30, e47.Google Scholar
  9. 9.
    Talmud, P., Tybjaeg-Hansen, A., Bhatnagar, D., et al. (1991) Rapid screening for specific mutations in patients with a clinical diagnosis of familial hypercholesterolaemia. Artherosclerosis 89, 137–141.CrossRefGoogle Scholar
  10. 10.
    Roche Molecular Biochemicals (2002) LightCycler Operators Manual version 3.5. Roche Diagnostics GmbH, Mannheim, Germany.Google Scholar

Copyright information

© Humana Press Inc 2004

Authors and Affiliations

  1. 1.Department of Molecular Medicine and HaematologyUniversity of the WitwatersrandJohannesburgSouth Africa
  2. 2.National Health Laboratory Services (NHLS)South Africa

Personalised recommendations