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Relative Human Telomere Length Quantification by Real-Time PCR

  • A. Vasilishina
  • A. Kropotov
  • I. Spivak
  • A. Bernadotte
Part of the Methods in Molecular Biology book series (MIMB, volume 1896)

Abstract

Telomere measurement by quantitative PCR amplification is a well-known simple method to detect telomere length that involves large numbers of samples. The method has been developed by Cawthon in 2002 (Cawthon, Nucleic Acids Res 30:47e–47, 2002) and remains the most frequently used technique either in original or modified version. Telomere length is estimated by comparing the amount of telomere repeat amplification product (T) to a single copy gene (S) product. The T/S ratio correlates with the average telomere length. Cawthon suggested and recommended the use of 36B4 (RPLP0) as a single copy gene. However, Cawthon’s suggestion was no longer considered a single copy gene and the gene was not suitable and appropriate for normalization.

We thereby introduced a simple method for relative measurement of average human telomere length using quantitative real-time PCR. Our protocol was based on Cawthon’s initial technique (Cawthon, Nucleic Acids Res 30:47e–47, 2002), modified by single-copy gene (SCG) primers and optimized.

This technique is rapid, low cost, not demanding on DNA amount (or live cells), and can be used for a high-throughput screening and time monitoring.

Key words

Telomere length Quantitative PCR Real-time PCR Telomere Telomere measurement IFNB1 36B4 Primers Single-copy gene 

References

  1. 1.
    Olovnikov A (1971) Principle of marginotomy in template synthesis of polynucleotides. Dokl Akad Nauk SSSR 201:1496–1499PubMedGoogle Scholar
  2. 2.
    Olovnikov A (1973) A theory of marginotomy. The incomplete copying of template margin in enzymic synthesis of polynucleotides and biological significance of the phenomenon. J Theor Biol 41:181–190CrossRefPubMedGoogle Scholar
  3. 3.
    Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30:47e.  https://doi.org/10.1093/nar/30.10.e47CrossRefGoogle Scholar
  4. 4.
    Callicott R, Womack J (2006) Real-time PCR assay for measurement of mouse telomeres. Comp Med 56:17–22PubMedGoogle Scholar
  5. 5.
    Wand T, Fang M, Chen C et al (2016) Telomere content measurement in human hematopoietic cells: comparative analysis of qPCR and Flow-FISH techniques. Cytometry 89:914–921CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • A. Vasilishina
    • 1
  • A. Kropotov
    • 1
  • I. Spivak
    • 1
  • A. Bernadotte
    • 2
    • 3
  1. 1.Institute of Cytology, Russian Academy of SciencesSaint-PetersburgRussian Federation
  2. 2.Faculty of Mechanics and MathematicsLomonosov Moscow State UniversityMoscowRussian Federation
  3. 3.Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Laboratory of Simple SystemsMoscowRussian Federation

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