How to statistically compare samples that only have 1 replicate


I am trying to understand if it is possible to compare statistically , differences amongst samples that only have 1 replicate…so basically there is no average!

In other words let’s say I have a otu table (.shared file) with 5 different samples and I want to compare if these 5 samples are statistically different to one another…

In this case my .design table would only have this 5 samples… Can this work when running an AMOVA?

Thank you!

With only one sample per group there is no way to calculate metrics like the mean and variation within a group that statistical test rely upon.

AMOVA specifically is a tricky one as it has minimum samples per group requirements that you can read about here. Because of the way the P-Values are calculated some combinations of numbers of samples per group will never reach significance.

With all that said, biological significance can still be derived from datasets that are too small to satisfy statistical significance.


With just 5 samples, you’re going to be limited to basically ordination to visually compare. And then describe the communities you found.

Ok Thanks!
I suspected it was really not possible…