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The majority of life on Earth is non-diploid and represented by prokaryotes, viruses and their derivatives such as our own mitochondria or plant’s chloroplasts. In non-diploid systems allele frequencies can range anywhere between 0 and 100% and there could be multiple (not just two) alleles per locus. The main challenge associated with non-diploid variant calling is the difficulty in distinguishing between sequencing noise (abundant in all NGS platforms) and true low frequency variants.