Real-time quantitative PCR assay for measurement of avain telomeres

Criscuolo, F., Bize, P., Nasir, L., Metcalfe, N.B., Foote, C.G., Griffiths, K., Gault, E.A. and Monaghan, P. (2009) Real-time quantitative PCR assay for measurement of avain telomeres. Journal of Avian Biology, 40(3), pp. 342-347. (doi:10.1111/j.1600-048X.2008.04623.x)

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Publisher's URL: http://dx.doi.org/10.1111/j.1600-048X.2008.04623.x

Abstract

We present the application of a real-time quantitative PCR assay, previously developed to measure relative telomere length in humans and mice, to two bird species, the zebra finch <i>Taeniopygia guttata</i> and the Alpine swift <i>Apus melba</i>. This technique is based on the PCR amplification of telomeric (TTAGGG)<sub>n</sub> sequences using specific oligonucleotide primers. Relative telomere length is expressed as the ratio (T/S) of telomere repeat copy number (T) to control single gene copy number (S). This method is particularly useful for comparisons of individuals within species, or where the same individuals are followed longitudinally. We used glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a single control gene. In both species, we validated our PCR measurements of relative telomere length against absolute measurements of telomere length determined by the conventional method of quantifying telomere terminal restriction fragment (TRF) lengths using both the traditional Southern blot analysis (Alpine swifts) and in gel hybridization (zebra finches). As found in humans and mice, telomere lengths in the same sample measured by TRF and PCR were well correlated in both the Alpine swift and the zebra finch.. Hence, this PCR assay for measurement of bird telomeres, which is fast and requires only small amounts of genomic DNA, should open new avenues in the study of environmental factors influencing variation in telomere length, and how this variation translates into variation in cellular and whole organism senescence

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Monaghan, Professor Patricia and Metcalfe, Professor Neil and Griffiths, Mrs Katharine and Gault, Mrs Elizabeth and Nasir, Professor Lubna and Bize, Dr Pierre and Criscuolo, Dr Francois
Authors: Criscuolo, F., Bize, P., Nasir, L., Metcalfe, N.B., Foote, C.G., Griffiths, K., Gault, E.A., and Monaghan, P.
Subjects:Q Science > QL Zoology
Q Science > QH Natural history > QH301 Biology
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
College of Medical Veterinary and Life Sciences > School of Veterinary Medicine
Journal Name:Journal of Avian Biology
ISSN:0908-8857

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
401931Life history consequences of growth accelerationPatricia MonaghanNatural Environment Research Council (NERC)NE/C004353/1RI BIODIVERSITY ANIMAL HEALTH & COMPMED