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>The antibiotic exposed group (N = 31) included 16 males (M) and 15 females (F), and the control group (N = 17) included nine males and eight females.

Small N but as a layman it could be potentially significant. Perhaps hormonal changes could be monitored in fecal microbiomes.

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Honestly, for PCR, this is a larger N than I would guess is needed. In general, rats kept in constant conditions don't have a ton of variation, even in fairly high-sensitivity assays. I would expect a lot more samples if you were sampling wild rats from the subway system.

I recently looked at a disease course over time for several hundred genes. I only used 6 animals per time point, and both the disease mice and non-disease mice have very little inter-mouse variation at each time point for the vast majority of the genes.

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