Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland.
Placenta. 2013 Jul;34(7):544-7. doi: 10.1016/j.placenta.2013.03.011. Epub 2013 Apr 25.
It has been highlighted that RNA quality and appropriate reference gene selection is crucial for the interpretation of RT-qPCR results in human placental samples. In this context we investigated the effect of RNA degradation on the mRNA abundance of seven frequently used reference genes in 119 human placental samples. Combining RNA integrity measurements, RT-qPCR analysis and mathematical modeling we found major differences regarding the effect of RNA degradation on the measured expression levels between the different reference genes. Furthermore, we demonstrated that a modified RNA extraction method significantly improved RNA quality and consequently increased transcript levels of all reference genes.
已有研究强调，在解读人胎盘样本 RT-qPCR 结果时，RNA 质量和适当的参考基因选择至关重要。在此背景下，我们研究了 RNA 降解对 119 个人胎盘样本中 7 个常用参考基因 mRNA 丰度的影响。通过 RNA 完整性测量、RT-qPCR 分析和数学建模，我们发现不同参考基因的 RNA 降解对测量表达水平的影响存在显著差异。此外，我们还证明了一种改良的 RNA 提取方法可显著改善 RNA 质量，从而提高所有参考基因的转录水平。
