Jiménez Karen M, Forero Diego A
Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, 110231, Colombia.
Mol Biol Rep. 2018 Aug;45(4):633-638. doi: 10.1007/s11033-018-4175-y. Epub 2018 Apr 5.
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.
端粒长度（TL）的改变与多种疾病相关，一种基于定量聚合酶链反应（qPCR）的方法，即单色多重实时定量PCR（MMQPCR）技术，已被广泛用于TL分析。先前的一些研究发现，某些方法学条件会影响TL的测量。本研究的目的是评估八种不同的市售SYBR Green和高分辨率熔解（HRM）混合试剂在通过MMQPCR方法测量TL方面的性能。测试了四种SYBR Green和四种HRM混合试剂，TL的测量用T/S比值表示。结果发现，MMQPCR中使用的预混试剂类型会影响TL的测量，影响结果的特异性和一致性等方面。我们的结果首次描述了不同预混试剂对MMQPCR分析TL的影响，并突出了未来改进这种广泛应用技术的方法学的重要性。
