Profiling Extracellular Vesicle miRNAs

The miRNA amount of the EV samples was calculated with the Qubit™ microRNA Assay Kit (Thermo Fisher Scientific, Germany). cDNA was synthesized using TaqMan^® MicroRNA Reverse Transcription Kit (Thermo Fisher Scientific, Germany). RT-PCR was performed using TaqMan^® universal PCR Master Mix (Thermo Fisher Scientific, Germany) on a Quantstudio three cycler (Applied Biosystems, Germany). The following miRNAs were examined: let7a, miR-10a, miR-21, miR-23, miR-155, miR-221, miR-222 and miR-486, miR29a, miR34a. All primers were purchased from Applied Biosystems (Supplementary Table S2). The samples were run in duplicates. U18 was used as endogenous control according to Vriens et al. (Vriens et al., 2012 (link)). Amplicons were normalized to U18 applying the comparative CT (∆CT) method.