Molecular Biology Reports

, Volume 45, Issue 4, pp 633–638 | Cite as

Effect of master mixes on the measurement of telomere length by qPCR

  • Karen M. Jiménez
  • Diego A. Forero
Short Communication


Alterations in telomere length (TL) have been associated with several diseases and a method based on qPCR, the Monochrome Multiplex Real-Time Quantitative PCR (MMQPCR) technique, has been used extensively for the analysis of TL. Some previous studies have been found that certain methodological conditions can affect the measurement of TL. The aim of the study was to evaluate the performance of eight different commercially available SYBR Green and High-Resolution Melting (HRM) mixes on the measurement of TL by the MMQPCR method. Four SYBR Green and four HRM mixes were tested and the measurement of TL was expressed by the T/S ratio. It was found that the type of master mix used in MMQPCR influences the measurement of TL, affecting aspects such as the specificity and consistency of the results. Our results are the first description of the effects of different master mixes on TL analysis by MMQPCR and highlight the importance of the future methodological improvement of this broadly used technique.


Epigenetics Molecular genetics Molecular methods qPCR Telomere length 



This study was supported by research grants from Colciencias (Grant No. 823-2015) and VCTI (Grant No. 2016220).

Compliance with ethical standards

Conflict of interest

The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in this study.


  1. 1.
    Chatterjee S (2017) Telomeres in health and disease. J Oral Maxillofac Pathol 21(1):87–91. CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Polho GB, De-Paula VJ, Cardillo G, dos Santos B, Kerr DS (2015) Leukocyte telomere length in patients with schizophrenia: a meta-analysis. Schizophr Res 165(2–3):195–200. CrossRefPubMedGoogle Scholar
  3. 3.
    Schutte NS, Malouff JM (2015) The association between depression and leukocyte telomere length: a meta-analysis. Depress Anxiety 32(4):229–238. CrossRefPubMedGoogle Scholar
  4. 4.
    Forero DA, Gonzalez-Giraldo Y, Lopez-Quintero C, Castro-Vega LJ, Barreto GE, Perry G (2016) Meta-analysis of telomere length in Alzheimer’s disease. J Gerontol Ser A 71(8):1069–1073. CrossRefGoogle Scholar
  5. 5.
    Malouff JM, Schutte NS (2017) A meta-analysis of the relationship between anxiety and telomere length. Anxiety Stress Coping 30(3):264–272. CrossRefPubMedGoogle Scholar
  6. 6.
    Ridout KK, Ridout SJ, Price LH, Sen S, Tyrka AR (2016) Depression and telomere length: a meta-analysis. J Affect Disord 191:237–247. CrossRefPubMedGoogle Scholar
  7. 7.
    Monroy-Jaramillo N, Dyukova E, Walss-Bass C (2017) Telomere length in psychiatric disorders: is it more than an ageing marker?. World J Biol Psychiatry. PubMedGoogle Scholar
  8. 8.
    Mathur MB, Epel E, Kind S, Desai M, Parks CG, Sandler DP, Khazeni N (2016) Perceived stress and telomere length: a systematic review, meta-analysis, and methodologic considerations for advancing the field. Brain Behav Immun 54:158–169. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Montpetit AJ, Alhareeri AA, Montpetit M, Starkweather AR, Elmore LW, Filler K, Mohanraj L, Burton CW, Menzies VS, Lyon DE, Jackson-Cook CK (2014) Telomere length: a review of methods for measurement. Nurs Res 63(4):289–299. CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Cawthon RM (2002) Telomere measurement by quantitative PCR. Nucleic Acids Res 30(10):e47CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Cawthon RM (2009) Telomere length measurement by a novel monochrome multiplex quantitative PCR method. Nucleic Acids Res 37(3):e21. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Hsieh AY, Saberi S, Ajaykumar A, Hukezalie K, Gadawski I, Sattha B, Cote HC (2016) Optimization of a relative telomere length assay by monochromatic multiplex real-time quantitative PCR on the LightCycler 480: sources of variability and quality control considerations. J Mol Diagn 18(3):425–437. CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Martin-Ruiz CM, Baird D, Roger L, Boukamp P, Krunic D, Cawthon R, Dokter MM, van der Harst P, Bekaert S, de Meyer T, Roos G, Svenson U, Codd V, Samani NJ, McGlynn L, Shiels PG, Pooley KA, Dunning AM, Cooper R, Wong A, Kingston A, von Zglinicki T (2015) Reproducibility of telomere length assessment: an international collaborative study. Int J Epidemiol 44(5):1673–1683. CrossRefPubMedGoogle Scholar
  14. 14.
    Eisenberg DT, Kuzawa CW, Hayes MG (2015) Improving qPCR telomere length assays: controlling for well position effects increases statistical power. Am J Hum Biol 27(4):570–575. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Denham J, Marques FZ, Charchar FJ (2014) Leukocyte telomere length variation due to DNA extraction method. BMC Res Notes 7:877. CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Forero DA, Gonzalez-Giraldo Y, Lopez-Quintero C, Castro-Vega LJ, Barreto GE, Perry G (2016) Telomere length in Parkinson’s disease: a meta-analysis. Exp Gerontol 75:53–55. CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Lahiri DK, Nurnberger JI Jr (1991) A rapid non-enzymatic method for the preparation of HMW DNA from blood for RFLP studies. Nucleic Acids Res 19(19):5444CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Cunningham JM, Johnson RA, Litzelman K, Skinner HG, Seo S, Engelman CD, Vanderboom RJ, Kimmel GW, Gangnon RE, Riegert-Johnson DL, Baron JA, Potter JD, Haile R, Buchanan DD, Jenkins MA, Rider DN, Thibodeau SN, Petersen GM, Boardman LA (2013) Telomere length varies by DNA extraction method: implications for epidemiologic research. Cancer Epidemiol Biomarker Prev 22(11):2047–2054. CrossRefGoogle Scholar
  19. 19.
    Zanet DL, Saberi S, Oliveira L, Sattha B, Gadawski I, Cote HC (2013) Blood and dried blood spot telomere length measurement by qPCR: assay considerations. PLoS ONE 8(2):e57787. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Yang J, Kemps-Mols B, Spruyt-Gerritse M, Anholts J, Claas F, Eikmans M (2016) The source of SYBR green master mix determines outcome of nucleic acid amplification reactions. BMC Res Notes 9:292. CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Eischeid AC (2011) SYTO dyes and EvaGreen outperform SYBR Green in real-time PCR. BMC Res Notes 4:263. CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Aubert G, Hills M, Lansdorp PM (2012) Telomere length measurement-caveats and a critical assessment of the available technologies and tools. Mutat Res 730(1–2):59–67. CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  1. 1.Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of MedicineUniversidad Antonio NariñoBogotáColombia

Personalised recommendations