Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia.
PhD Program in Health Sciences, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia.
Biotechniques. 2019 Oct;67(4):192-199. doi: 10.2144/btn-2019-0065. Epub 2019 Sep 6.
Several approaches for miRNA expression analysis have been developed in recent years. In this article, we provide an updated and comprehensive review of available qPCR-based methods for miRNA expression analysis and discuss their advantages and disadvantages. Existing techniques involve the use of stem-loop reverse transcriptase-PCR, polyadenylation of RNAs, ligation of adapters or RT with complex primers, using universal or miRNA-specific qPCR primers and/or probes. Many of these methods are oriented towards the expression analysis of mature miRNAs and few are designed for the study of pre-miRNAs and pri-miRNAs. We also discuss findings from articles that compare results from existing methods. Finally, we suggest key points for the improvement of available techniques and for the future development of additional methods.
近年来，已经开发出了几种 miRNA 表达分析的方法。在本文中，我们提供了基于 qPCR 的 miRNA 表达分析方法的最新和全面的综述，并讨论了它们的优缺点。现有的技术涉及使用茎环逆转录酶-PCR、RNA 的多聚腺苷酸化、接头的连接或带有复杂引物的 RT、使用通用或 miRNA 特异性 qPCR 引物和/或探针。这些方法中的许多方法都针对成熟 miRNA 的表达分析，而很少针对 pre-miRNA 和 pri-miRNA 的研究设计。我们还讨论了比较现有方法结果的文章中的发现。最后，我们提出了改进现有技术和开发其他方法的要点。
