Hello,
I have been working with plasma miRNAs for a long time and yesterday I obtained some very strange results, which I had never seen before. As you can see in the attached photo, there is a strong but decreasing fluorescence signal detected from the beginning of the qPCR, before there is any amplification. Secondly, some of the curves are normal until a certain point and either stop before the end of the procedure, or continue to increase in an abnormal pattern. Could someone explain these findings?
Do you think there might be a primer dimers' problem?
Would it help if I diluted more the cDNA (I diluted it 12 times and there is nothing appearing when I put just water)?
Thank you in advance for your help.
Regards,
Nikiforos
Plasma microRNAs should not be so abundant, in humans they are mostly >30 Cts. The amplification curve on this do not look normal (should be more steep?). Do you have a NTC? It would be informative to see the signal coming from that.
Several plasma miRNAs are particularly abundant (miR-16, miR-21, miR-451) and generally detected at
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