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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
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.
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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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