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

TY - JOUR

T1 - Bio-On-Magnetic-Beads (BOMB)

T2 - Open platform for high-throughput nucleic acid extraction and manipulation

AU - Oberacker, Phil

AU - Stepper, Peter

AU - Bond, Donna M

AU - Höhn, Sven

AU - Focken, Jule

AU - Meyer, Vivien

AU - Schelle, Luca

AU - Sugrue, Victoria J

AU - Jeunen, Gert-Jan

AU - Moser, Tim

AU - Hore, Steven R

AU - von Meyenn, Ferdinand

AU - Hipp, Katharina

AU - Hore, Timothy A

AU - Jurkowski, Tomasz P

PY - 2019/1/10

Y1 - 2019/1/10

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85060393925&partnerID=8YFLogxK

U2 - 10.1371/journal.pbio.3000107

DO - 10.1371/journal.pbio.3000107

M3 - Article

C2 - 30629605

SN - 1544-9173

VL - 17

SP - e3000107

JO - PLoS Biology

JF - PLoS Biology

IS - 1

M1 - e3000107

ER -