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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...

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Autores principales: Selz, Jonathan, Adam, Nicolas R., Magrini, Céline E. M., Montandon, Fulvia Malvido, Buerki, Sven, Maerkl, Sebastian J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2023
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.
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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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