Methodology for Extracting High-Molecular-Weight DNA from Field Collections of Macrofungi

Many macrofungi are impractical or impossible to culture. Consequently, DNA for long-read sequencing required for the assembly of high-quality genomes must be isolated from samples taken from the environment. Collection is often in remote locations, limiting the options for stabilising samples to methods that do not require refrigeration. Fungi contain species-specific arrays of metabolites that may complicate purification techniques and call for judgement to be made to apply appropriate modifications to the DNA extraction protocol in specific cases. The protocols and commentary we describe are informed by the preparation of DNA from a range of Australasian ectomycorrhizal and saprotrophic macrofungi. We collect samples into isopropanol at ambient temperature and employ a strategy of chromatin isolation followed by the sequential removal of unwanted molecular components to purify DNA.

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Publication Details

Journal
Journal of Fungi
Published
2025-06-27
DOI
https://doi.org/10.3390/jof11070490
Primary Topic
Mycorrhizal Fungi and Plant Interactions
Type
article
Field-Weighted Citation Impact
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article

Methodology for Extracting High-Molecular-Weight DNA from Field Collections of Macrofungi

Vivian Ng, Igor V. Grigoriev, David Catcheside, Tom W. May et al.
Journal of Fungi
Mycorrhizal Fungi and Plant Interactions
article

Methodology for Extracting High-Molecular-Weight DNA from Field Collections of Macrofungi

Vivian Ng, Igor V. Grigoriev, David Catcheside, Tom W. May, Teresa Lebel, Anna Lipzen, Pamela S. Catcheside, David A. Orlovich, Alan Kuo, Robert Riley, Maxim Koriabine, Andy R. Nilsen, William Andreopoulos, Kurt LaButti, Mi Yan, Leigh A. Burgoyne
article en

Abstract

Many macrofungi are impractical or impossible to culture. Consequently, DNA for long-read sequencing required for the assembly of high-quality genomes must be isolated from samples taken from the environment. Collection is often in remote locations, limiting the options for stabilising samples to methods that do not require refrigeration. Fungi contain species-specific arrays of metabolites that may complicate purification techniques and call for judgement to be made to apply appropriate modifications to the DNA extraction protocol in specific cases. The protocols and commentary we describe are informed by the preparation of DNA from a range of Australasian ectomycorrhizal and saprotrophic macrofungi. We collect samples into isopropanol at ambient temperature and employ a strategy of chromatin isolation followed by the sequential removal of unwanted molecular components to purify DNA.

Journal of FungiVol. 11(7)
Lawrence Berkeley National Laboratory (US), Flinders University (AU), Joint Genome Institute (US), State Herbarium of South Australia (AU), Royal Botanic Garden Sydney (AU), Royal Botanic Gardens Victoria, University of Otago (NZ), University of California, Berkeley (US)
Openalex Percentile: Top 89%
Mycorrhizal Fungi and Plant Interactions
DOIOpen Access PDFOpenAlex
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