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a quantitative pcr method for measuring absolute telomere length绝对定量pcr方法测量端粒长度
O’Callaghan and Fenech Biological Procedures Online 2011, 13:3
/content/13/1/3 Biological Procedures
Online
METHODOLOGY Open Access
A quantitative PCR method for measuring
absolute telomere length
*
Nathan J O’Callaghan , Michael Fenech
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.
Introduction decline, as well as total survival independent of genetic
Telomeres are nucleoprotein structures that cap the ends influences [18-24].
of chromosomes. The integrity of the telomere structure For all of these reasons there has been a burgeoning
and its DNA hexamer (TTAGGG)n re
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