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Biological Procedures Online

, 13:3

First Online: 31 January 2011Received: 07 October 2010Accepted: 31 January 2011DOI: 10.1186-1480-9222-13-3

Cite this article as: O-Callaghan, N.J. & Fenech, M. Biol Proced Online 2011 13: 3. doi:10.1186-1480-9222-13-3

Abstract

We describe a simple and reproducible method to measure absolute telomere length aTL using quantitative real-time polymerase chain reaction qPCR. This method is based on the Cawthon method for relative measurement of telomere length TL but modified by introducing an oligomer standard to measure aTL. The method describes the oligomer standards, the generation of the standard curve and the calculations required to calculate aTL from the qPCR data. The necessary controls and performance characteristics of the assay are described in detail and compared relative to other methods for measuring TL. Typical results for this assay for a variety of human tissue samples are provided as well as a troubleshooting schedule. This method allows high throughput measurement of aTL using small amounts of DNA making it amenable for molecular epidemiological studies. Compared to the traditional relative TL qPCR assays, the aTL method described in this protocol enables a more direct comparison of results between experiments within and between laboratories.

Electronic supplementary materialThe online version of this article doi:10.1186-1480-9222-13-3 contains supplementary material, which is available to authorized users.

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Autor: Nathan J O-Callaghan - Michael Fenech

Fuente: https://link.springer.com/







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