Real-time quantitative PCR of telomere length
Purchase on Springer.com
$39.95 / €34.95 / £29.95*
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.
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.
- Cech, T. R. (2000) Life at the end of the chromosome: telomeres and telomerase. Angew. Chem. Int. Ed. Engl. 39, 34–43. CrossRef
- Hahn, W. C. (2003) Role of telomeres and telomerase in the pathogenesis of human cancer. J. Clin. Oncol. 21, 2034–2043. CrossRef
- Mathon, N. F. and Lloyd, A. C. (2001) Cell senescence and cancer. Nat. Rev. Cancer 1, 203–213. CrossRef
- Harley, C. B., Futcher, A. B., and Greider, C. W. (1990) Telomeres shorten during ageing of human fibroblasts. Nature 345, 458–460. CrossRef
- 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.
- 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.
- 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. CrossRef
- Cawthon, R. M. (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res. 30, e47.
- 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. CrossRef
- Roche Molecular Biochemicals (2002) LightCycler Operators Manual version 3.5. Roche Diagnostics GmbH, Mannheim, Germany.
- Real-time quantitative PCR of telomere length
Volume 27, Issue 2 , pp 169-172
- Cover Date
- Print ISSN
- Online ISSN
- Humana Press
- Additional Links
- telomere length
- real-time PCR
- Industry Sectors