Missing data in vcf output format about snp-sites

[ViriatoII]

I've very interested in knowing the answer to this. No one helps?

[BiologicalScientist]

I have also noticed this. I assume it is due to the fact that N is the ambiguity code for any base so there isn't strictly a variant in that location. It would be nice to have the option to call strictly anything that differs from the reference even if there is ambiguity or missing data. Not sure if the other ambiguity codes are similarly called the same

As a workaround for this I believe you could change all Ns in your fasta file to something that doesn't have a IUPAC meaning like a J or means missing data which is typically . or - That won't be confused for another base or amino acid (should you be using those). Just make sure to change it back in your output. Worth noting for an amino acid case you might find there to be more ambiguity codes that are an issue (though I haven't tried to see if Y and R etc also cause issues.)