Simple Synthesis of Functionalized Paramagnetic Beads for Nucleic Acid Purification and Manipulation

Phil Oberacker; Peter Stepper; Donna Bond; Katharina Hipp; Timothy A Hore; Tomasz P Jurkowski

doi:10.21769/BioProtoc.3394

Simple Synthesis of Functionalized Paramagnetic Beads for Nucleic Acid Purification and Manipulation

Bio Protoc. 2019 Oct 20;9(20):e3394. doi: 10.21769/BioProtoc.3394.

Authors

Phil Oberacker^{1

2}, Peter Stepper¹, Donna Bond³, Katharina Hipp⁴, Timothy A Hore³, Tomasz P Jurkowski²

Affiliations

¹ Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
² School of Biosciences, Cardiff University, Cardiff, Wales, United Kingdom.
³ Department of Anatomy, University of Otago, Dunedin, New Zealand.
⁴ Max Planck Institute for Developmental Biology, Tübingen, Germany.

Abstract

The purification of nucleic acids is one of the most common procedures employed in modern molecular biology laboratories. Typically, commercial column-based protocols are utilized to isolate DNA or RNA from various sources. However, these methods not only require specialized equipment, but are also extremely expensive for high-throughput applications. Although an elegant answer to this issue can be provided by paramagnetic beads, bead-based open-source protocols have been limited in the past. Here, we provide an easy to follow step-by-step manual for the synthesis of paramagnetic beads, as well as their functionalization with either a silica- or a carboxyl-surface that can be used to replace the commercial columns with self-made magnetic beads. Together with a variety of detailed protocols for their use in high-throughput nucleic acids extractions, this bead synthesis method forms the recently published open platform Bio-On-Magnetic-Beads (BOMB), which is available on PLOS Biology ( Oberacker et al., 2019 ). Updated protocols can be found on the associated webpage (https://bomb.bio).

Keywords: Fe3O4; SiO2; Bead synthesis; Carboxyl-coating; Magnetic beads; Magnetic nanoparticles; Magnetic separation; Methacrylic acid; Silica-coating; Surface functionalization.