With @benoitevol.bsky.social, we developed our own strategy to properly estimate π (e.g. doi.org/10.1016/j.cu...) and I extended it to variable ploidy (doi.org/10.1093/gene...). But it's clearly not as user-friendly & computationally efficient as pixy for similar results ➡️Use pixy, forget VCFtools!😉
😍 pixy 😍 - "[...] implementations of new estimators of Watterson's θ and Tajima's D that are unbiased with respect to missing data. - [...] support for arbitrary ploidy and multiallelic sites. - [...] optimizations, including multicore execution" REPEAT AFTER ME: There is no reason not to use Pixy.