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Assessment of isomiR Discrimination Using Commercial qPCR Methods.

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.

SUBMITTER: Magee R

PROVIDER: S-EPMC5514785 | biostudies-literature | 2017 Jun

REPOSITORIES: biostudies-literature

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Assessment of isomiR Discrimination Using Commercial qPCR Methods.

Magee Rogan R Telonis Aristeidis G AG Cherlin Tess T Rigoutsos Isidore I Londin Eric E

Non-coding RNA 20170324 2

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 infl ...[more]

PMID: 28730153

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