Evaluation of three RT-qPCR-based miRNA detection methods using seven rice miRNAs.

Three frequently-used reverse transcription-quantitative polymerase chain reaction (RT-qPCR)-based miRNA detection methods, stem-loop RT-qPCR, poly(A)-tailing RT-qPCR, and miQPCR, were evaluated using seven selected rice miRNAs. The results revealed that miRNA abundance and sequence characteristics can affect capability of detection. The stem-loop amplification technique detected highly and moderately abundant miRNAs. The poly(A)-tailing method detected both highly abundant and sparsely present miRNAs, but failed to detect miRNAs with a hairpin structure. Only a few miRNAs were detectable by the miQPCR method. We suggest that a combination of methods should be used for reliable quantitative investigation of miRNAs.