Hello everyone,
I've been doing circulating miRNA qPCR. Since there is not a generally acknowledged endogenous reference, we used cel-miR-39-3p as an external control. However, we found that the raw Ct values of cel-miR-39 varies from sample to sample, it seems even not as stable as hsa-miR-16, which is used as an alternative control (though there are some problems). The median and CV of raw Ct values for cel-miR-39 and hsa-miR-16 are 16 and 24, 12% and 6%, respectively.
I used Qiagen miRNeasy kit for extraction, TaqMan RT kit and TaqMan probe for qPCR. All RNA were extracted from 170ul corresponding plasma, diluted in 60ul RNase-free water.
Looking for your help, thanks very much!
Juncheng
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