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Uninterruptible Power Supply Improves Precision of Telomere Length Measurement via qPCR

机译:不间断电源通过QPCR提高了端粒长度测量的精度

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摘要

Epidemiological literature has produced robust associations between telomere length (TL) and health, wherein individuals with shorter TL are at increased risk for chronic disease and death. Even so, technical challenges associated with TL measurement have led some to question their utility as a biomarker of aging. Several pre-analytical factors influence TL assessment via qPCR including tissue source, sample storage, and DNA extraction method. Additional work has investigated how conditions within the PCR run (e.g. mastermix reagents) influences precision and reproducibility of TL measurements. However, the impact of power supply remains unclear. Momentary fluctuations in power supply can affect the functioning of high-performance electronics, including real-time thermocylers. These fluctuations can be mitigated by using an uninterruptible power source (UPS), an electronic apparatus capable of supplying constant, uninterrupted voltage to high-performance electronics. The current study investigated how using a UPS influenced TL assessment via qPCR. Standard deviation and coefficient of variation across replicates were compared for samples assessed with or without the use of a UPS. Samples run with a UPS had significantly lower standard deviation (p<0.001) and coefficient of variation (p=0.002) than those run without a UPS. Notably, neither the efficiency of exponential amplification (p=0.674) nor the standard curve R2 (p=0.638) varied as a function of UPS status. Thus, UPS status decreases variability within sample replicates while maintaining the overall quality of the qPCR assay. *The work presented was supported by the Telomere Research Network, an NIH-sponsored working-group recently established to coordinate best practices for measuring telomere length in population-based research.
机译:流行病学文献在端粒长度(TL)和健康之间产生了鲁棒的关联,其中TL短TL的个体患慢性疾病和死亡的风险增加。即便如此,与TL测量相关的技术挑战导致一些问题作为衰老的生物标志物质疑他们的效用。几种预分析因素通过QPCR影响TL评估,包括组织源,样品储存和DNA提取方法。另外的工作研究了PCR运行中的情况(例如MasterMix试剂)如何影响T1测量的精度和再现性。但是,电源的影响仍不清楚。电源的瞬间波动会影响高性能电子器件的功能,包括实时热膜。可以通过使用不间断电源(UPS),能够向高性能电子设备提供恒定的不间断电压的电子设备来缓解这些波动。目前的研究调查了如何通过QPCR使用UPS影响TL评估。比较随着使用或不使用UPS评估的样品对复制的标准偏差和变异系数。使用UPS运行的样本显着降低标准偏差(P <0.001)和变化系数(P = 0.002),而不是没有UPS的那些。值得注意的是,指数放大的效率(P = 0.674)和标准曲线R2(P = 0.638)无与伦比的UPS状态的函数变化。因此,UPS状态降低了样品复制的可变性,同时保持QPCR测定的整体质量。 *呈现的工作是由Teleromere研究网络支持的,该组是最近建立的NIH赞助的工作组,以协调衡量基于人群的研究的最佳实践。

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