Assessment of isomiR Discrimination Using Commercial qPCR Methods.

Noncoding RNA

Computational Medicine Center, http://cm.jefferson.edu/, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA 19107, USA.

Published: June 2017


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Article Abstract

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.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5514785PMC
http://dx.doi.org/10.3390/ncrna3020018DOI Listing

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