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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.
Afiliación
  • Oberacker P; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Stepper P; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Bond DM; Department of Anatomy, University of Otago, Dunedin, New Zealand.
  • Höhn S; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Focken J; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Meyer V; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Schelle L; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
  • Sugrue VJ; Department of Anatomy, University of Otago, Dunedin, New Zealand.
  • Jeunen GJ; Department of Anatomy, University of Otago, Dunedin, New Zealand.
  • Moser T; Department of Anatomy, University of Otago, Dunedin, New Zealand.
  • Hore SR; CENENG Ltd, Omakau, New Zealand.
  • von Meyenn F; Epigenetics Programme, Babraham Institute, Cambridge, United Kingdom and Department of Medical and Molecular Medicine, Kings College London, Guys Hospital, London, United Kingdom.
  • Hipp K; Max Planck Institute for Developmental Biology, Tübingen, Germany.
  • Hore TA; Department of Anatomy, University of Otago, Dunedin, New Zealand.
  • Jurkowski TP; Department of Biochemistry, Institute of Biochemistry and Technical Biochemistry, University of Stuttgart, Stuttgart, Germany.
PLoS Biol ; 17(1): e3000107, 2019 01.
Article en En | MEDLINE | ID: mdl-30629605
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.
Asunto(s)

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Ácidos Nucleicos / Secuenciación de Nucleótidos de Alto Rendimiento Tipo de estudio: Guideline Idioma: En Revista: PLoS Biol Asunto de la revista: BIOLOGIA Año: 2019 Tipo del documento: Article

Texto completo: 1 Base de datos: MEDLINE Asunto principal: Ácidos Nucleicos / Secuenciación de Nucleótidos de Alto Rendimiento Tipo de estudio: Guideline Idioma: En Revista: PLoS Biol Asunto de la revista: BIOLOGIA Año: 2019 Tipo del documento: Article