Extraction and Purification of DNA from Wood at Various Stages of Decay for Metabarcoding of Wood-Associated Fungi

Jeff R Powell; Michaela Blyton; Brad Oberle; Gillian L Powell; Jessica Rigg; Darcy Young; Amy E Zanne

doi:10.1007/978-1-0716-1040-4_10

Extraction and Purification of DNA from Wood at Various Stages of Decay for Metabarcoding of Wood-Associated Fungi

Methods Mol Biol. 2021:2232:113-122. doi: 10.1007/978-1-0716-1040-4_10.

Authors

Jeff R Powell¹, Michaela Blyton^{2

3}, Brad Oberle^{4

5}, Gillian L Powell², Jessica Rigg^{2

6}, Darcy Young⁵, Amy E Zanne⁵

Affiliations

¹ Hawkesbury Institute for the Environment, Western Sydney University, Penrith, NSW, Australia. jeff.powell@westernsydney.edu.au.
² Hawkesbury Institute for the Environment, Western Sydney University, Penrith, NSW, Australia.
³ School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, Australia.
⁴ Division of Natural Sciences, New College of Florida, Sarasota, FL, USA.
⁵ Department of Biological Science, George Washington University, Washington, DC, USA.
⁶ NSW Department of Primary Industries, Elizabeth Macarthur Agricultural Institute, Menangle, NSW, Australia.

PMID: 33161543
DOI: 10.1007/978-1-0716-1040-4_10

Abstract

Assessment of endophytic and saprotrophic microbial communities from wood-extracted DNA presents challenges due to the presence of surface microbes that contaminate samples and plant compounds that act as inhibiting agents. Here, we describe a method for decontaminating, sampling, and processing wood at various stages of decay for high-throughput extraction and purification of DNA.

Keywords: CTAB; Comparative analysis; Decomposition; Shavings; Stems; Surface decontamination.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

DNA / genetics
DNA / isolation & purification*
DNA Barcoding, Taxonomic / methods*
Fungi / classification
Fungi / genetics*
Wood / genetics*
Wood / microbiology

Substances

DNA