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Invariant sites not included in no_sites
Hello,
I am trying to use pixy on data generated using a reduced representation RAD protocol. I assembled loci using the Stacks program (Catchen et al. 2013) and generated a vcf including both variant and invariant sites (data_subset.vcf.gz) with the Stacks Populations module. However, when I run pixy on the dataset, the results indicate that only the variant loci are considered valid and used for calculating pi. In the attached data and output, chromosome (RAD locus) 4 contains 303 total sites, 7 of which are variant. However, in the pixy_pi_subset.txt output file, you can see that no_sites = 7, suggesting that only 7 sites "have at least one valid genotype", according to the Pixy documentation. What is causing my invariant sites be considered invalid genotypes?
Code used to run Pixy:
pixy --stats pi fst dxy --vcf data_subset.vcf.gz --populations popmap.txt --window_size 10000 --n_cores 4
data_subset.vcf.gz (note: needs to be indexed prior to running as .tbi file would not attach) popmap.txt pixy_pi.txt
Thank you, Geoffrey