I order real-time PCR , cDNA kit and 8 primers from exiqon. but under the exiqon protocol, I couldn't see any results for one primer ( miR-625) The company says that the expression level of this miRNA was so low that kit can not detect it .
Hi
Some miRNAs are present at very low expression levels. I would advise to increase the amplification cycles in qPCR (45-50) to see if you can detect something. Check with the melting curve for primer dimers or inespecific amplification.
Best
If you're concerned that something might be wrong with that particular assay then I'd recommend you use a RNA sample that is know to express miR-625.
I agree with Jeffrey. You should perform a new PCR using a true positive control (some kind of sample that is known to express a great amount of miR-625) to test the primer viability.
I think that the primer has the problem and miRNA expression isn't so low (by considering the attached papers .
As seen in attached picture by adding 2 micrograms of primer the CT of 38 was obtained .
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