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Ancient DNA extraction methods for herbarium specimens: When is it worth the effort?
PREMISE: Herbaria harbor a tremendous number of plant specimens that are rarely used for molecular systematic studies, largely due to the difficulty in extracting sufficient amounts of high‐quality DNA from the preserved plant material. METHODS: We compared the standard Qiagen DNeasy Plant Mini Kit...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9215277/ https://www.ncbi.nlm.nih.gov/pubmed/35774991 http://dx.doi.org/10.1002/aps3.11477 |
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author | Marinček, Pia Wagner, Natascha D. Tomasello, Salvatore |
author_facet | Marinček, Pia Wagner, Natascha D. Tomasello, Salvatore |
author_sort | Marinček, Pia |
collection | PubMed |
description | PREMISE: Herbaria harbor a tremendous number of plant specimens that are rarely used for molecular systematic studies, largely due to the difficulty in extracting sufficient amounts of high‐quality DNA from the preserved plant material. METHODS: We compared the standard Qiagen DNeasy Plant Mini Kit and a specific protocol for extracting ancient DNA (aDNA) (the N‐phenacylthiazolium bromide and dithiothreitol [PTB–DTT] extraction method) from two different plant genera (Xanthium and Salix). The included herbarium materials covered about two centuries of plant collections. To analyze the success of DNA extraction using each method, a subset of samples was subjected to a standard library preparation as well as target‐enrichment approaches. RESULTS: The PTB–DTT method produced a higher DNA yield of better quality than the Qiagen kit; however, extracts from the Qiagen kit over a certain DNA yield and quality threshold produced comparable sequencing results. The sequencing resulted in high proportions of endogenous reads. We were able to successfully sequence 200‐year‐old samples. DISCUSSION: This method comparison revealed that, for younger specimens, DNA extraction using a standard kit might be sufficient. For old and precious herbarium specimens, aDNA extraction methods are better suited to meet the requirements for next‐generation sequencing. |
format | Online Article Text |
id | pubmed-9215277 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-92152772022-06-29 Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? Marinček, Pia Wagner, Natascha D. Tomasello, Salvatore Appl Plant Sci Application Article PREMISE: Herbaria harbor a tremendous number of plant specimens that are rarely used for molecular systematic studies, largely due to the difficulty in extracting sufficient amounts of high‐quality DNA from the preserved plant material. METHODS: We compared the standard Qiagen DNeasy Plant Mini Kit and a specific protocol for extracting ancient DNA (aDNA) (the N‐phenacylthiazolium bromide and dithiothreitol [PTB–DTT] extraction method) from two different plant genera (Xanthium and Salix). The included herbarium materials covered about two centuries of plant collections. To analyze the success of DNA extraction using each method, a subset of samples was subjected to a standard library preparation as well as target‐enrichment approaches. RESULTS: The PTB–DTT method produced a higher DNA yield of better quality than the Qiagen kit; however, extracts from the Qiagen kit over a certain DNA yield and quality threshold produced comparable sequencing results. The sequencing resulted in high proportions of endogenous reads. We were able to successfully sequence 200‐year‐old samples. DISCUSSION: This method comparison revealed that, for younger specimens, DNA extraction using a standard kit might be sufficient. For old and precious herbarium specimens, aDNA extraction methods are better suited to meet the requirements for next‐generation sequencing. John Wiley and Sons Inc. 2022-06-15 /pmc/articles/PMC9215277/ /pubmed/35774991 http://dx.doi.org/10.1002/aps3.11477 Text en © 2022 The Authors. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of Botanical Society of America. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Application Article Marinček, Pia Wagner, Natascha D. Tomasello, Salvatore Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title | Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title_full | Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title_fullStr | Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title_full_unstemmed | Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title_short | Ancient DNA extraction methods for herbarium specimens: When is it worth the effort? |
title_sort | ancient dna extraction methods for herbarium specimens: when is it worth the effort? |
topic | Application Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9215277/ https://www.ncbi.nlm.nih.gov/pubmed/35774991 http://dx.doi.org/10.1002/aps3.11477 |
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