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Evaluation of gene expression by the "2–∆∆Ct" method.

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Are you rows here technical replicates or biological replicates? Normally we should calculate the average ct across our technical replicates. And then calculate separately a ddct for each of the biological replicates (using the average of the technical replicates).

Numbers from 1 to 5 or from 11 to 15 mean different organisms (i.e. 5 different animals in the control and experimental group and the only difference between them is a dietary supplement). And during real-time PCR, I did three repetitions for each organism. cDNA from one organism (for example, number 1) got into the wells of the PCR tablet A1-B1-C1 and then I made an average of the values.

As far as I understand, these are biological replicates.

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