To date, a significant amount of work has been performed on the human microbiome to evaluate its composition and influence on physiology; this research has led to additional studies on microbiomes localized at specific sites of the human body (e.g., skin, oral, vaginal). Given that fungi are ubiquitous and live in symbiosis with the human body, researchers are now actively looking into the role of the mycobiome in human health and disease. Recent advancements in sequencing technologies have enabled the community profiling of fungi; however, the complexities associated with metagenomics sequencing analyses have posed significant challenges toward standardization. To address this need, ATCC has developed genomic DNA and whole cell mock microbial communities comprising ten medically relevant fungal species mixed in even proportions. In this proof-of-concept study, we demonstrate the use these standards in evaluating DNA extraction and sequencing methods for mycobiome analysis.