Real-time PCR assay for measurement of mouse telomeres.
Measurement of telomeres by polymerase chain reaction (PCR) amplification has been problematic due to the formation of dimers by the primers designed to hybridize to the telomere repeats. Recently, a set of primers that overcome this problem has been created and used to develop an assay to measure human telomeres by real-time quantitative PCR. We modified this assay to measure mouse telomeres. Results showed that the primers do indeed amplify mammalian telomere repeats without forming dimers. Results obtained from the real-time quantitative PCR assay of mouse DNA were similar to terminal restriction fragment analysis by pulsed-field gel electrophoresis followed by Southern hybridization. The assay performed with mouse DNA in a similar manner as it performs with human DNA. Preliminary linkage mapping suggests a gene influencing telomere length on the X chromosome. This assay will aid in the study of telomere function and importance in diseases associated with aging and cancer formation.
Callicott
Ralph J
RJ
Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University College Station, Texas, USA. rcallicott@cvm.tamu.edu
Womack
James E
JE
D000818:Animals; D002874:Chromosome Mapping; D017931:DNA Primers / Q000235:genetics; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D013045:Species Specificity; D016615:Telomere / Q000235:genetics* / Q000648:ultrastructure
D017931:DNA Primers