烟曲霉高通量培养及DNA分离方法

Current protocols Pub Date : 2025-04-02 DOI:10.1002/cpz1.70112

Francisca C. Reyes Márquez, Ben Auxier, Bo Briggeman, Eveline Snelders

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引用次数: 0

摘要

烟曲霉是一种在环境中广泛传播的机会性人类真菌病原体。在使用目前可用的方案时，需要筛选大量的环境或临床分离株以进行表型和/或基因型目的，这可能是具有挑战性和耗时的。我们已经开发了一种新的方法，允许人们接种培养物，培养单个烟曲霉分离物，并随后分离出足够质量的DNA进行PCR和/或全基因组测序，所有这些都在96孔格式。与目前可用的方法相比，我们的方案大大降低了成本和人工，并且与任何使用96井格式的实验装置兼容。©2025作者。Wiley期刊有限责任公司发表的当前方案：基本方案1：烟曲霉的96孔微型斜培养和孢子悬浮液的制备；基本方案2:pcr质量DNA的分离；基本方案3：微型菌丝板培养后高纯度DNA的分离

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High-Throughput Culture and DNA Isolation Methods for Aspergillus fumigatus

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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

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来源期刊