hi, everyone. I am detecting mature microRNA expression level these days using taqman probe in vivo system. I want to know whether I can still trust data when the Cq value of microRNAs reach 30?
Yes, don't worry this is very classical. I had the same concern when I began doing qPCR for miRNAs and several experts in the field told me that this was normal. The cut-off is not the same as for classical genes.
The only way to figure out if you can trust your data in this case is to use the many controls that can be used for a qPCR including (for the most basic ones) assessment of replicates and consistency of the dissociation curve.
Dear Julien,
How did you use control for microrna detection? Also, I am not sure whether I use the right protocol for RT. I have to detect 12 microRNAs. I did reverse transcription separately, and the reference gene is reverse transcribed alone. Could you please give me a detailed protocol for reverse transcription
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