in analyzing qPCR results, I saw the Ct value for one of my miRNAs is less than the two different selected ref. genes in the patient's condition, but not in control samples.
Should I change my ref. genes?
For calculating ΔCт , ΔCт value is positive value always?in any condition?
Is it possible the patient's condition cause this overexpression of this miRNA?
what about your negative controls? if no signal at the same Ct range,then you are fine.
Additionally, if the technical replicates are constant, then again you are fine, even if your internal control genes shows lower Ct values.
Dear Ahmed
I have no signal at the same Ct range in my negative controls.
The Ct values for control groups are around 20, but in patient samples around 18 in all 3 replicates
and
The Ct for my two reference genes are around 19.
Dear Carin
Sincerely Thank you for your guidance
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