Currently I’m working on an experiment for evaluation of IL-17 (interlukin-17) expression, I used house-keeping gene (GAPDH) as an endogenous control. I started with RNA and used a single step RT-PCR mix and a realtime PCR. Now I’ve the Cts and not quite sure what to do with the analysis. I don’t have a standard curve for this gene, but I have another one. I’ve been advised to use relative quantitation (ΔCt). But I’m not familiar with the steps of calculating or how I’m going to consider this gene expressed or not. Also there’re some software tools for this matter does anyone here have one?
Thanks for your help