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https://hdl.handle.net/2440/126163
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Type: | Journal article |
Title: | An optimized method for the extraction of ancient eukaryote DNA from marine sediments |
Author: | Armbrecht, L. Herrando-Pérez, S. Eisenhofer, R. Hallegraeff, G.M. Bolch, C.J.S. Cooper, A. |
Citation: | Molecular Ecology Resources, 2020; 20(4):906-919 |
Publisher: | Wiley |
Issue Date: | 2020 |
ISSN: | 1755-098X 1755-0998 |
Statement of Responsibility: | Linda Armbrecht, Salvador Herrando-Pérez, Raphael Eisenhofer, Gustaaf M. Hallegraeff, Christopher J. S. Bolch, Alan Cooper |
Abstract: | Marine sedimentary ancient DNA (sedaDNA) provides a powerful means to reconstruct marine palaeo-communities across the food web. However, currently there are few optimized sedaDNA extraction protocols available to maximize the yield of small DNA fragments typical of ancient DNA (aDNA) across a broad diversity of eukaryotes. We compared seven combinations of sedaDNA extraction treatments and sequencing library preparations using marine sediments collected at a water depth of 104 m off Maria Island, Tasmania, in 2018. These seven methods contrasted frozen versus refrigerated sediment, bead-beating induced cell lysis versus ethylenediaminetetraacetic acid (EDTA) incubation, DNA binding in silica spin columns versus in silica-solution, diluted versus undiluted DNA in shotgun library preparations to test potential inhibition issues during amplification steps, and size-selection of low molecular-weight (LMW) DNA to increase the extraction efficiency of sedaDNA. Maximum efficiency was obtained from frozen sediments subjected to a combination of EDTA incubation and bead-beating, DNA binding in silica-solution, and undiluted DNA in shotgun libraries, across 45 marine eukaryotic taxa. We present an optimized extraction protocol integrating these steps, with an optional post-library LMW size-selection step to retain DNA fragments of ≤500 base pairs. We also describe a stringent bioinformatic filtering approach for metagenomic data and provide a comprehensive list of contaminants as a reference for future sedaDNA studies. The new extraction and data-processing protocol should improve quantitative paleo-monitoring of eukaryotes from marine sediments, as well as other studies relying on the detection of highly fragmented and degraded eukaryote DNA in sediments. |
Keywords: | ancient DNA diatoms dinoflagellates haptophytes Maria Island metagenomics plankton seafloor Tasmania |
Rights: | © 2020 John Wiley & Sons Ltd |
DOI: | 10.1111/1755-0998.13162 |
Grant ID: | http://purl.org/au-research/grants/arc/CE170100015 http://purl.org/au-research/grants/arc/DP170102261 http://purl.org/au-research/grants/arc/FL140100260 |
Published version: | http://dx.doi.org/10.1111/1755-0998.13162 |
Appears in Collections: | Aurora harvest 8 Australian Centre for Ancient DNA publications |
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