I have read about various ways to measure the gene abundance ratio, Pfaffl, deltadelta Ct, etc. I want to measure the change in target gene abundance at 3 different timepoints. I have the Ct values for all three timepoints per subject and then I did a 16S pcr with the same samples used for the three timepoints as my reference gene. How could i best analyse the change in the gene abundance while normalizing for my 16S?