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Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids
BACKGROUND: Plant DNA isolation and purification is a time-consuming and laborious process relative to epithelial and viral DNA sample preparation due to the cell wall. The lysis of plant cells to free intracellular DNA normally requires high temperatures, chemical surfactants, and mechanical separa...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8943943/ https://www.ncbi.nlm.nih.gov/pubmed/35321738 http://dx.doi.org/10.1186/s13007-022-00860-8 |
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author | Emaus, Miranda N. Cagliero, Cecilia Gostel, Morgan R. Johnson, Gabriel Anderson, Jared L. |
author_facet | Emaus, Miranda N. Cagliero, Cecilia Gostel, Morgan R. Johnson, Gabriel Anderson, Jared L. |
author_sort | Emaus, Miranda N. |
collection | PubMed |
description | BACKGROUND: Plant DNA isolation and purification is a time-consuming and laborious process relative to epithelial and viral DNA sample preparation due to the cell wall. The lysis of plant cells to free intracellular DNA normally requires high temperatures, chemical surfactants, and mechanical separation of plant tissue prior to a DNA purification step. Traditional DNA purification methods also do not aid themselves towards fieldwork due to the numerous chemical and bulky equipment requirements. RESULTS: In this study, intact plant tissue was coated by hydrophobic magnetic ionic liquids (MILs) and ionic liquids (ILs) and allowed to incubate under static conditions or dispersed in a suspension buffer to facilitate cell disruption and DNA extraction. The DNA-enriched MIL or IL was successfully integrated into the qPCR buffer without inhibiting the reaction. The two aforementioned advantages of ILs and MILs allow plant DNA sample preparation to occur in one minute or less without the aid of elevated temperatures or chemical surfactants that typically inhibit enzymatic amplification methods. MIL or IL-coated plant tissue could be successfully integrated into a qPCR assay without the need for custom enzymes or manual DNA isolation/purification steps that are required for conventional methods. CONCLUSIONS: The limited amount of equipment, chemicals, and time required to disrupt plant cells while simultaneously extracting DNA using MILs makes the described procedure ideal for fieldwork and lab work in low resource environments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-022-00860-8. |
format | Online Article Text |
id | pubmed-8943943 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-89439432022-03-25 Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids Emaus, Miranda N. Cagliero, Cecilia Gostel, Morgan R. Johnson, Gabriel Anderson, Jared L. Plant Methods Research BACKGROUND: Plant DNA isolation and purification is a time-consuming and laborious process relative to epithelial and viral DNA sample preparation due to the cell wall. The lysis of plant cells to free intracellular DNA normally requires high temperatures, chemical surfactants, and mechanical separation of plant tissue prior to a DNA purification step. Traditional DNA purification methods also do not aid themselves towards fieldwork due to the numerous chemical and bulky equipment requirements. RESULTS: In this study, intact plant tissue was coated by hydrophobic magnetic ionic liquids (MILs) and ionic liquids (ILs) and allowed to incubate under static conditions or dispersed in a suspension buffer to facilitate cell disruption and DNA extraction. The DNA-enriched MIL or IL was successfully integrated into the qPCR buffer without inhibiting the reaction. The two aforementioned advantages of ILs and MILs allow plant DNA sample preparation to occur in one minute or less without the aid of elevated temperatures or chemical surfactants that typically inhibit enzymatic amplification methods. MIL or IL-coated plant tissue could be successfully integrated into a qPCR assay without the need for custom enzymes or manual DNA isolation/purification steps that are required for conventional methods. CONCLUSIONS: The limited amount of equipment, chemicals, and time required to disrupt plant cells while simultaneously extracting DNA using MILs makes the described procedure ideal for fieldwork and lab work in low resource environments. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-022-00860-8. BioMed Central 2022-03-24 /pmc/articles/PMC8943943/ /pubmed/35321738 http://dx.doi.org/10.1186/s13007-022-00860-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Emaus, Miranda N. Cagliero, Cecilia Gostel, Morgan R. Johnson, Gabriel Anderson, Jared L. Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title | Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title_full | Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title_fullStr | Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title_full_unstemmed | Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title_short | Simple and efficient isolation of plant genomic DNA using magnetic ionic liquids |
title_sort | simple and efficient isolation of plant genomic dna using magnetic ionic liquids |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8943943/ https://www.ncbi.nlm.nih.gov/pubmed/35321738 http://dx.doi.org/10.1186/s13007-022-00860-8 |
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