Reverse transcription for miRNAs was performed with a qScript flex cDNA synthesis kit (Quantabio, Beverly, MA, USA) according to Androvic et al. [47 (link)] in the total reaction volume of 10 µL. RNA was diluted in TE-LPA buffer (TE buffer with linear polyacrylamide at a final working concentration of 20 µg/mL). The RT reaction mixture contained 10 ng of total RNA, 1 × RT buffer, 0.05 µM RT primer, 1 µL GSP enhancer, and 0.5 µL RT enzyme. RT reactions were incubated in PCR tubes for 45 min at 25 °C and for 5 min at 85 °C and then held at 4 °C.
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