I'm analysis miRNA expression in serum samles. I have 30 controls and 50 asthmatics. How can I calculate delta delta Ct value? There is no control matched to each study sample, as you can see. Can I use 1 sample from a healthy group as a CALIBRATOR and based on it calculate delta delta Ct for the rest of samples (also the rest of samples from the healthy group)? I also did calculation with the mean delta Ct of all healthy individuals and I obtained similar final results. But is it correct way of relative quantification?
Dear Malgorzata,
If you've earned similar final results for healthy individuals, and then you've used average, in my opinion your way is true.
Good luck,
Hi ... u have used endogenous microRNA or house keeping such as miR -16 or any other depending on reference paper
Then please check this reference to do ur calculations
Good luck
Ye the way that you took one of the control samples as calibrator sample is the way relative quantification works.
Relative quantification is always based on fold change in expression of gene under study in ratios of control and treatment samples... so your work is on track.. best wishes
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