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High-Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus

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Open Access
Type
Article
Language
en
Date
2025-04-02
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Title
High-Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus
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Curr protocol 2025; 5(4):e70112
Abstract
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.
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