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Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation
Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge- and column-based protocols require specialised equipment, often use toxic reagents, and are not economically scalable or practical to use in a high-throughput man...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6343928/ https://www.ncbi.nlm.nih.gov/pubmed/30629605 http://dx.doi.org/10.1371/journal.pbio.3000107 |
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author | Oberacker, Phil Stepper, Peter Bond, Donna M. Höhn, Sven Focken, Jule Meyer, Vivien Schelle, Luca Sugrue, Victoria J. Jeunen, Gert-Jan Moser, Tim Hore, Steven R. von Meyenn, Ferdinand Hipp, Katharina Hore, Timothy A. Jurkowski, Tomasz P. |
author_facet | Oberacker, Phil Stepper, Peter Bond, Donna M. Höhn, Sven Focken, Jule Meyer, Vivien Schelle, Luca Sugrue, Victoria J. Jeunen, Gert-Jan Moser, Tim Hore, Steven R. von Meyenn, Ferdinand Hipp, Katharina Hore, Timothy A. Jurkowski, Tomasz P. |
author_sort | Oberacker, Phil |
collection | PubMed |
description | Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge- and column-based protocols require specialised equipment, often use toxic reagents, and are not economically scalable or practical to use in a high-throughput manner. Although it has been known for some time that magnetic beads can provide an elegant answer to these issues, the development of open-source protocols based on beads has been limited. In this article, we provide step-by-step instructions for an easy synthesis of functionalised magnetic beads, and detailed protocols for their use in the high-throughput purification of plasmids, genomic DNA, RNA and total nucleic acid (TNA) from a range of bacterial, animal, plant, environmental and synthetic sources. We also provide a bead-based protocol for bisulfite conversion and size selection of DNA and RNA fragments. Comparison to other methods highlights the capability, versatility, and extreme cost-effectiveness of using magnetic beads. These open-source protocols and the associated webpage (https://bomb.bio) can serve as a platform for further protocol customisation and community engagement. |
format | Online Article Text |
id | pubmed-6343928 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-63439282019-02-01 Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation Oberacker, Phil Stepper, Peter Bond, Donna M. Höhn, Sven Focken, Jule Meyer, Vivien Schelle, Luca Sugrue, Victoria J. Jeunen, Gert-Jan Moser, Tim Hore, Steven R. von Meyenn, Ferdinand Hipp, Katharina Hore, Timothy A. Jurkowski, Tomasz P. PLoS Biol Community Page Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge- and column-based protocols require specialised equipment, often use toxic reagents, and are not economically scalable or practical to use in a high-throughput manner. Although it has been known for some time that magnetic beads can provide an elegant answer to these issues, the development of open-source protocols based on beads has been limited. In this article, we provide step-by-step instructions for an easy synthesis of functionalised magnetic beads, and detailed protocols for their use in the high-throughput purification of plasmids, genomic DNA, RNA and total nucleic acid (TNA) from a range of bacterial, animal, plant, environmental and synthetic sources. We also provide a bead-based protocol for bisulfite conversion and size selection of DNA and RNA fragments. Comparison to other methods highlights the capability, versatility, and extreme cost-effectiveness of using magnetic beads. These open-source protocols and the associated webpage (https://bomb.bio) can serve as a platform for further protocol customisation and community engagement. Public Library of Science 2019-01-10 /pmc/articles/PMC6343928/ /pubmed/30629605 http://dx.doi.org/10.1371/journal.pbio.3000107 Text en © 2019 Oberacker et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Community Page Oberacker, Phil Stepper, Peter Bond, Donna M. Höhn, Sven Focken, Jule Meyer, Vivien Schelle, Luca Sugrue, Victoria J. Jeunen, Gert-Jan Moser, Tim Hore, Steven R. von Meyenn, Ferdinand Hipp, Katharina Hore, Timothy A. Jurkowski, Tomasz P. Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title | Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title_full | Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title_fullStr | Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title_full_unstemmed | Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title_short | Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation |
title_sort | bio-on-magnetic-beads (bomb): open platform for high-throughput nucleic acid extraction and manipulation |
topic | Community Page |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6343928/ https://www.ncbi.nlm.nih.gov/pubmed/30629605 http://dx.doi.org/10.1371/journal.pbio.3000107 |
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