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PLoS Biol. 2019 Jan 10;17(1):e3000107. doi: 10.1371/journal.pbio.3000107. [Epub ahead of print]

Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation.

Author information

1
Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
2
Department of Anatomy, University of Otago, Dunedin, New Zealand.
3
CENENG Ltd, Omakau, New Zealand.
4
Epigenetics Programme, Babraham Institute, Cambridge, United Kingdom and Department of Medical and Molecular Medicine, Kings College London, Guys Hospital, London, United Kingdom.
5
Max Planck Institute for Developmental Biology, Tübingen, Germany.

Abstract

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.

PMID:
30629605
DOI:
10.1371/journal.pbio.3000107
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Conflict of interest statement

TH and DB are directors of a small agricultural and biotech consultancy (TOTOGEN Ltd), at which some BOMB protocols were developed and have been donated to this project. SRH is director of an agricultural engineering firm (CENENG Ltd).

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