So far, surveys of the mycobiome have been hampered by the lack of fungal marker genes that are as universal as prokaryotic 16S rRNA genes. The internal transcribed spacer 1 (ITS1) region in the eukaryotic ribosomal cluster has been used as a marker gene, but the currently available primers have a limited potential to recover fungal diversity and are biased towards soil fungi. Usyk et al. have now developed a new set of ITS1 primers, which almost double the taxonomic coverage of a reference database and substantially increase the number of recovered sequences from fungal isolates compared with previously published primers. Notably, the coverage of the Candida genus, which includes important human pathogens, increased from ∼60% to 100%. Increased taxonomic coverage and sequencing depth should facilitate future mycobiome studies.