Magee Rogan, Telonis Aristeidis G, Cherlin Tess, Rigoutsos Isidore, Londin Eric
Computational Medicine Center, http://cm.jefferson.edu/, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA 19107, USA.
Noncoding RNA. 2017 Jun;3(2). doi: 10.3390/ncrna3020018. Epub 2017 Mar 24.
We sought to determine whether commercial quantitative polymerase chain reaction (qPCR) methods are capable of distinguishing isomiRs: variants of mature microRNAs (miRNAs) with sequence endpoint differences. We used two commercially available miRNA qPCR methods to quantify miR-21-5p in both synthetic and real cell contexts. We find that although these miRNA qPCR methods possess high sensitivity for specific sequences, they also pick up background signals from closely related isomiRs, which influences the reliable quantification of individual isomiRs. We conclude that these methods do not possess the requisite specificity for reliable isomiR quantification.
我们试图确定商业定量聚合酶链反应（qPCR）方法是否能够区分同型微小RNA（isomiRs）：即成熟微小RNA（miRNAs）具有序列端点差异的变体。我们使用两种市售的miRNA qPCR方法在合成和真实细胞环境中对miR-21-5p进行定量。我们发现，尽管这些miRNA qPCR方法对特定序列具有高灵敏度，但它们也会从密切相关的同型微小RNA中检测到背景信号，这会影响单个同型微小RNA的可靠定量。我们得出结论，这些方法不具备可靠定量同型微小RNA所需的特异性。
