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A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring
PREMISE: A novel protocol for rapid plant DNA extraction using microneedles is proposed, which supports botanic surveys, taxonomy, and systematics. This protocol can be conducted in the field with limited laboratory skills and equipment. The protocol is validated by sequencing and comparing the resu...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10278934/ https://www.ncbi.nlm.nih.gov/pubmed/37342168 http://dx.doi.org/10.1002/aps3.11529 |
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author | Selz, Jonathan Adam, Nicolas R. Magrini, Céline E. M. Montandon, Fulvia Malvido Buerki, Sven Maerkl, Sebastian J. |
author_facet | Selz, Jonathan Adam, Nicolas R. Magrini, Céline E. M. Montandon, Fulvia Malvido Buerki, Sven Maerkl, Sebastian J. |
author_sort | Selz, Jonathan |
collection | PubMed |
description | PREMISE: A novel protocol for rapid plant DNA extraction using microneedles is proposed, which supports botanic surveys, taxonomy, and systematics. This protocol can be conducted in the field with limited laboratory skills and equipment. The protocol is validated by sequencing and comparing the results with QIAGEN spin‐column DNA extractions using BLAST analyses. METHODS AND RESULTS: Two sets of DNA extractions were conducted on 13 species spanning various leaf anatomies and phylogenetic lineages: (i) fresh leaves were punched with custom polymeric microneedle patches to recover genomic DNA, or (ii) QIAGEN DNA extractions. Three plastid (matK, rbcL, and trnH‐psbA) and one nuclear ribosomal (ITS) DNA regions were amplified and sequenced using Sanger or nanopore technology. The proposed method reduced the extraction time to 1 min and yielded the same DNA sequences as the QIAGEN extractions. CONCLUSIONS: Our drastically faster and simpler method is compatible with nanopore sequencing and is suitable for multiple applications, including high‐throughput DNA‐based species identifications and monitoring. |
format | Online Article Text |
id | pubmed-10278934 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-102789342023-06-20 A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring Selz, Jonathan Adam, Nicolas R. Magrini, Céline E. M. Montandon, Fulvia Malvido Buerki, Sven Maerkl, Sebastian J. Appl Plant Sci Protocol Note PREMISE: A novel protocol for rapid plant DNA extraction using microneedles is proposed, which supports botanic surveys, taxonomy, and systematics. This protocol can be conducted in the field with limited laboratory skills and equipment. The protocol is validated by sequencing and comparing the results with QIAGEN spin‐column DNA extractions using BLAST analyses. METHODS AND RESULTS: Two sets of DNA extractions were conducted on 13 species spanning various leaf anatomies and phylogenetic lineages: (i) fresh leaves were punched with custom polymeric microneedle patches to recover genomic DNA, or (ii) QIAGEN DNA extractions. Three plastid (matK, rbcL, and trnH‐psbA) and one nuclear ribosomal (ITS) DNA regions were amplified and sequenced using Sanger or nanopore technology. The proposed method reduced the extraction time to 1 min and yielded the same DNA sequences as the QIAGEN extractions. CONCLUSIONS: Our drastically faster and simpler method is compatible with nanopore sequencing and is suitable for multiple applications, including high‐throughput DNA‐based species identifications and monitoring. John Wiley and Sons Inc. 2023-06-14 /pmc/articles/PMC10278934/ /pubmed/37342168 http://dx.doi.org/10.1002/aps3.11529 Text en © 2023 The Authors. Applications in Plant Sciences published by Wiley Periodicals LLC on behalf of 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 Selz, Jonathan Adam, Nicolas R. Magrini, Céline E. M. Montandon, Fulvia Malvido Buerki, Sven Maerkl, Sebastian J. A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title | A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title_full | A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title_fullStr | A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title_full_unstemmed | A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title_short | A field‐capable rapid plant DNA extraction protocol using microneedle patches for botanical surveying and monitoring |
title_sort | field‐capable rapid plant dna extraction protocol using microneedle patches for botanical surveying and monitoring |
topic | Protocol Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10278934/ https://www.ncbi.nlm.nih.gov/pubmed/37342168 http://dx.doi.org/10.1002/aps3.11529 |
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