Evaluation of Three RT-qPCR-Based miRNA Detection Methods Using Seven Rice miRNAs

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  • MOU Guiping
    College of Natural Resources and the Environment, South China Agricultural University
  • WANG Kang
    College of Natural Resources and the Environment, South China Agricultural University
  • XU Donglin
    College of Natural Resources and the Environment, South China Agricultural University
  • ZHOU Guohui
    College of Natural Resources and the Environment, South China Agricultural University

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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.

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