High-Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus

Aspergillus fumigatus is an opportunistic human fungal pathogen that is widely spread in the environment. The need to screen a large number of environmental or clinical isolates for phenotypic and/or genotypic purposes can be challenging and time consuming when using the protocols that are currently available. We have developed a novel approach that allows one to inoculate cultures, grow individual A. fumigatus isolates, and subsequently isolate DNA of sufficient quality for PCR and/or whole genome sequencing, all in a 96-well format. Compared to currently available methods, our protocols reduce both cost and labor significantly and are compatible with any experimental setup that uses a 96-well format. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: 96-well-format mini slant culture and preparation of spore suspension for Aspergillus fumigatus Basic Protocol 2: PCR-quality DNA isolation Basic Protocol 3: High-purity DNA isolation after mini mycelial mat culture.