Hello,

I was asked to perform a standard curve process using qPCR after isolation of miRNAs from serum and plasma exosomes. I normally use the Fragment analyzer to estimate the concentration of total RNA present in my samples and then perform a qPCR to estimate the Ct values. For the serum and plasma samples, there are no established endogenous controls because of which I was asked to perform the qPCR to validate the accurate concentrations of my samples. I normally use spike-in ath-miR-159a during the total RNA isolation to enhance the extraction efficiency, but have never performed the standard curve process using qPCR. Can someone explain how I could do this? We make use of the TaqMan Advanced assays for the qPCR. 

Thanks,