Quantifying myomiR-206 expression using qPCR

Reverse transcription reactions for myomiR‐206 and U6 small nuclear RNA (Rnu6) were performed with 10 ng of total RNA using Taqman MicroRNA Reverse Transcription Kit (ThermoFisher Scientific) according to the manufacturer's directions. qPCR was carried out with Taqman Gene expression Master Mix (2×) (ThermoFisher Scientific), TaqMan gene expression assay (miR‐206, #000510; Rnu6, #001973) using cDNA in a 10 µl reaction volume. qPCR was performed using the QuantStudio3 (Applied Biosystems) qPCR system as described by the manufacturer. qPCR efficiency was calculated by linear regression during the exponential phase using LinRegPCR software v11.126 (Ruijter et al., 2009 (link)). The comparison of miRNAs expression between groups was determined following normalization with Rnu6. Relative quantification of miRNA expression was assessed by the ΔΔCT method relative to the control.