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A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa
PREMISE: Commonly used molecular techniques such as next‐generation sequencing require reliable methods to extract DNA quickly and efficiently. Secondary compounds within plant tissues make this requirement all the more challenging, often forcing researchers to test different extraction methods tail...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8027365/ https://www.ncbi.nlm.nih.gov/pubmed/33854847 http://dx.doi.org/10.1002/aps3.11413 |
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author | Mavrodiev, Evgeny V. Dervinis, Christopher Whitten, William Mark Gitzendanner, Matthew A. Kirst, Matias Kim, Sangtae Kinser, Taliesin J. Soltis, Pamela S. Soltis, Douglas E. |
author_facet | Mavrodiev, Evgeny V. Dervinis, Christopher Whitten, William Mark Gitzendanner, Matthew A. Kirst, Matias Kim, Sangtae Kinser, Taliesin J. Soltis, Pamela S. Soltis, Douglas E. |
author_sort | Mavrodiev, Evgeny V. |
collection | PubMed |
description | PREMISE: Commonly used molecular techniques such as next‐generation sequencing require reliable methods to extract DNA quickly and efficiently. Secondary compounds within plant tissues make this requirement all the more challenging, often forcing researchers to test different extraction methods tailored to their particular species of interest in order to obtain large amounts of high‐quality genomic DNA. The opportunities provided by high‐throughput, next‐generation sequencing only exacerbate these problems, especially when trying to extract DNA from multiple species at the same time. Several methods have attempted to resolve the challenges of obtaining suitable DNA from plants; however, a rapid, high‐yield, high‐quality, and highly scalable DNA extraction method is still needed. METHODS AND RESULTS: We present a rapid DNA extraction protocol that utilizes a buffer with relatively large amounts of cetyltrimethylammonium bromide (CTAB) and sodium chloride, combined with a silica maxi‐column cleanup of the extracted DNA. The new method is easy to implement using standard equipment and inexpensive reagents. The entire procedure (from grinding to the final elution) can be completed in less than two hours for a single sample and can be easily scaled to meet desired research goals. It works on diverse green plants with highly varied secondary chemistries (e.g., ferns, gymnosperms, and phylogenetically divergent angiosperms). CONCLUSIONS: Application of the protocol to various plant species yielded DNA of high quality in less than two hours and can be adjusted to extract DNA at large (maxi‐preps) or small (96‐well minipreps) scales. We anticipate that our method will be of wide utility for rapidly isolating large quantities of quality genomic DNA from diverse plant species and will have broad applications in phylogenetic studies utilizing PCR and short‐read DNA sequencing. |
format | Online Article Text |
id | pubmed-8027365 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-80273652021-04-13 A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa Mavrodiev, Evgeny V. Dervinis, Christopher Whitten, William Mark Gitzendanner, Matthew A. Kirst, Matias Kim, Sangtae Kinser, Taliesin J. Soltis, Pamela S. Soltis, Douglas E. Appl Plant Sci Protocol Note PREMISE: Commonly used molecular techniques such as next‐generation sequencing require reliable methods to extract DNA quickly and efficiently. Secondary compounds within plant tissues make this requirement all the more challenging, often forcing researchers to test different extraction methods tailored to their particular species of interest in order to obtain large amounts of high‐quality genomic DNA. The opportunities provided by high‐throughput, next‐generation sequencing only exacerbate these problems, especially when trying to extract DNA from multiple species at the same time. Several methods have attempted to resolve the challenges of obtaining suitable DNA from plants; however, a rapid, high‐yield, high‐quality, and highly scalable DNA extraction method is still needed. METHODS AND RESULTS: We present a rapid DNA extraction protocol that utilizes a buffer with relatively large amounts of cetyltrimethylammonium bromide (CTAB) and sodium chloride, combined with a silica maxi‐column cleanup of the extracted DNA. The new method is easy to implement using standard equipment and inexpensive reagents. The entire procedure (from grinding to the final elution) can be completed in less than two hours for a single sample and can be easily scaled to meet desired research goals. It works on diverse green plants with highly varied secondary chemistries (e.g., ferns, gymnosperms, and phylogenetically divergent angiosperms). CONCLUSIONS: Application of the protocol to various plant species yielded DNA of high quality in less than two hours and can be adjusted to extract DNA at large (maxi‐preps) or small (96‐well minipreps) scales. We anticipate that our method will be of wide utility for rapidly isolating large quantities of quality genomic DNA from diverse plant species and will have broad applications in phylogenetic studies utilizing PCR and short‐read DNA sequencing. John Wiley and Sons Inc. 2021-04-07 /pmc/articles/PMC8027365/ /pubmed/33854847 http://dx.doi.org/10.1002/aps3.11413 Text en © 2021 Mavrodiev et al. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of the Botanical Society of America https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Protocol Note Mavrodiev, Evgeny V. Dervinis, Christopher Whitten, William Mark Gitzendanner, Matthew A. Kirst, Matias Kim, Sangtae Kinser, Taliesin J. Soltis, Pamela S. Soltis, Douglas E. A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title | A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title_full | A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title_fullStr | A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title_full_unstemmed | A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title_short | A new, simple, highly scalable, and efficient protocol for genomic DNA extraction from diverse plant taxa |
title_sort | new, simple, highly scalable, and efficient protocol for genomic dna extraction from diverse plant taxa |
topic | Protocol Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8027365/ https://www.ncbi.nlm.nih.gov/pubmed/33854847 http://dx.doi.org/10.1002/aps3.11413 |
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