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Comparative Study
. 2011 Nov 1;39(20):e134.
doi: 10.1093/nar/gkr634. Epub 2011 Aug 8.

Impartial comparative analysis of measurement of leukocyte telomere length/DNA content by Southern blots and qPCR

Abraham Aviv  1 , Steven C Hunt, Jue Lin, Xiaojian Cao, Masayuki Kimura, Elizabeth Blackburn
Affiliations
Comparative Study

Impartial comparative analysis of measurement of leukocyte telomere length/DNA content by Southern blots and qPCR

Abraham Aviv et al. Nucleic Acids Res. .

Abstract

Telomere length/DNA content has been measured in epidemiological/clinical settings with the goal of testing a host of hypotheses related to the biology of human aging, but often the conclusions of these studies have been inconsistent. These inconsistencies may stem from various reasons, including the use of different telomere length measurement techniques. Here, we report the first impartial evaluation of measurements of leukocyte telomere length by Southern blot of the terminal restriction fragments and quantitative PCR (qPCR) of telomere DNA content, expressed as the ratio of telomeric product (T)/single copy gene (S) product. Blind measurements on the same samples from 50 donors were performed in two independent laboratories on two different occasions. Both the qPCR and Southern blots displayed highly reproducible results as shown by r values > 0.9 for the correlations between results obtained by either method on two occasions. The inter-assay CV measurement for the qPCR was 6.45%, while that of the Southern blots was 1.74%. The relation between the results generated by Southern blots versus those generated by qPCR deviated from linearity. We discuss the ramifications of these findings with regard to measurements of telomere length/DNA content in epidemiological/clinical circumstances.

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Figures

Figure 1.
Figure 1.
Correlations between the first set of LTL measurements and second set of measurements by qPCR (A) and Southern blot analysis (B) and the inter-assay variation ±95% confidence interval of the two methods (C).
Figure 2.
Figure 2.
Linear (A) and curvilinear (B) models depicting the relation between the mean TRF length and the T/S ratio.
Figure 3.
Figure 3.
Cross sectional analysis of age-dependent telomere shortening based on the qPCR (A) and Southern blots (B).
See this image and copyright information in PMC

References

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