Assessment of isomiR Discrimination Using Commercial qPCR Methods
Assessment of isomiR Discrimination Using Commercial qPCR Methods
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DOI：
10.3390/ncrna3020018
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发表时间：
2017-06-01
期刊：
影响因子：
4.3
通讯作者：
Londin, Eric
中科院分区：
文献类型：
--
作者：
Magee, Rogan;Telonis, Aristeidis G.;Londin, Eric
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isomiRs：具有序列终点差异的成熟microRNA（miRNAs）变体。我们使用两种市售的miRNA qPCR方法来定量合成和真实的细胞环境中的miR-21- 5 p。我们发现，尽管这些miRNA qPCR方法对特定序列具有高灵敏度，但它们也从密切相关的isomiR中提取背景信号，这影响了个体isomiR的可靠定量。我们得出结论，这些方法不具备可靠的isomiR定量所需的特异性。
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.