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 mann...

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Published inbioRxiv
Main Authors Oberacker, Phil, Stepper, Peter, Bond, Donna M, Hohn, Sven, Focken, Jule, Meyer, Vivien, Schelle, Luca, Sugrue, Victoria J, Gert-Jan Jeunen, Moser, Tim, Hore, Steven R, Ferdinand Von Meyenn, Hipp, Katharina, Hore, Timothy A, Jurkowski, Tomasz P
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 12.09.2018
Subjects
Bisulfite
Deoxyribonucleic acid
DNA
Plasmids
Purification
Ribonucleic acid
RNA
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Summary: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 and total RNA from different sources, as well as environmental TNA and PCR amplicons. 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.
DOI:10.1101/414516