Hi!I need help to analyze some Real Time PCR data. I want to measure the fold change of the gene X, using ACTI as constitutive expressed gene.

The point is: I need to make a technical duplicate and to work in triplicate (so for each gene I will have 6 different values). How should I proceed? I need to evaluate the ΔCt: should I evaluate the average of all the 6 values? Or should I first consider the average of the technical duplicates and then make the averege of the three? I'm asking this because if that's the case, I don't know how to handle the error (we generally use standard deviation)

I hope it's clear, I'm very confused...

EDIT:
https://www.youtube.com/watch?v=Kkle8T7aXjk&ab_channel=TopTipBio
I would do it this way, but again, how do I evaluate the error?