I performed 4 indipend qPCR runs in which controls and treated samples are tested for a goi with the relative ref gene.
each run represents a biological replicate
For each replicate I evaluate the fold change using the 2^-ddct method
is it possible to perform any statistical analysis as the t-test on these values if the control is setted at 1 and the variance is zero?
No, because you do not have "raw data values" to compare. You should perform the t-test at the ΔCt stage of the Ct data transformation to fold change as at this stage you still have comparable values for both control and treated that can be tested statistically.
Thanks Christyn.
I though that was possible to compare four independent fold changes from fuor replicates. The fold differences between control and treated samples are maintained along different experiments. But the same is not true for dct values in which the differences are less evident due to the relative high SD. Do you have another suggestion?
You can use the one-sample t-test with the log fold-changes.
Another option is a paired-sample t-test with the ΔCt values.
The results will be identical.
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