miRNA profiling
Serumexosomal RNAsamples isolatedwith UC (n=5) or EQ (n=5)
methods from healthy women were used for miRNA profiling by real
time PCR using miRCURY LNA™ microRNA Human panel I assays
(Exiqon, Vedbaek, Denmark) following the manufacturer's recommendations.
Briefly, cDNA synthesis was performed using the Universal
cDNA Synthesis Kit (Exiqon). For each reaction 4 μL of exosomal RNA
was used. Reverse transcription reaction products were combined
with SYBR® Green master mix (Exiqon) and loaded onto ready-to-use
Human Panel assays. Amplification was performed on a 7900HT
thermocycler (Applied Biosystems) using cycling parameters recommended
by Exiqon.