@inbook{095b1c664f034aad9afbaceb5bb27763,
title = "Tissue Cultivation, Preparation, and Extraction of High Molecular Weight DNA for Single-Molecule Genome Sequencing of Plant-Associated Fungi",
abstract = "Extraction of high-quality, high molecular weight DNA is a critical step for sequencing an organism{\textquoteright}s genome. For fungi, DNA extraction is often complicated by co-precipitation of secondary metabolites, the most destructive being polysaccharides, polyphenols, and melanin. Different DNA extraction protocols and clean-up methods have been developed to address challenging materials and contaminants; however, the method of fungal cultivation and tissue preparation also plays a critical role to limit the production of inhibitory compounds prior to extraction. Here, we provide protocols and guidelines for (i) fungal tissue cultivation and processing with solid media containing a cellophane overlay or in liquid media, (ii) DNA extraction with customized recommendations for taxonomically and ecologically diverse plant-associated fungi, and (iii) assessing DNA quantity and quality for downstream genome sequencing with single-molecule technology such as PacBio.",
keywords = "Cellophane overlay, Culturing methods, DNA extraction, DNA quality control, Fruiting bodies, Fungal growth, Liquid culture, Secondary metabolites",
author = "Laure Fauchery and Maxim Koriabine and Moore, {Lillian P.} and Yuko Yoshinaga and Kerrie Barry and Annegret Kohler and U{\textquoteright}Ren, {Jana M.}",
note = "Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2023",
doi = "10.1007/978-1-0716-2871-3_5",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "79--102",
booktitle = "Methods in Molecular Biology",
}