Welcome everyone!
In my research I focus on comparing Staphylococcus aureus genes to the reference gene - gmk. In qPCR I’m using relative quantification and the following formula:
∆Ct (target)= Ct sample - Ct control
∆Ct (calibrator)= Ct sample - Ct control
∆∆Ct= ∆Ct (target) - ∆Ct(calibrator)
R=2-∆∆Ct
Calibrator is my cells which were not treated. My first question is - is calibrator should have standard value or it depends, because I saw the information, that calibrator should not exceed 1.
Moreover, I have a problem with interpretation of R value. More of my results gave values below the 1 (eg. 0.40; 0.70). How I should interpreted these results? Is it correct if I say, that the expression level of my target gene is 40-fold or 70-fold lower than the expression in the reference gene?
Thank you in advance for your help!
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