My question is if is necessary to check normal distribution for perfoming Ancom or Aldex2 in metagenomic data, that is, a table of relative abundances of microbial species obtained by 16s or shotgun sequencing. As far as I know, ALDEx2 performs Welch’s t-test that requires normal distribution.
And in case it is necessary to check that distribution, with what free program can it be done easily?
The short answer is no. The compositional assumption and transformation under Aldex2 is more important than the assumption of normality.
Under the hood, Aldex2 uses a specific data transform with an underlying monte carlo simulation to achieve that transformation. Som if you wanted to test normality, you would need to approach the transformed data.