Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation
| Autor: | Phil Oberacker, Gert-Jan Jeunen, Jule Focken, Sven Hohn, Steven R. Hore, Luca Schelle, Timothy A. Hore, Peter Stepper, Donna M. Bond, Tomasz P. Jurkowski, Katharina Hipp, Vivien Meyer, Tim Moser, Victoria J Sugrue, Ferdinand von Meyenn |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Epidemiology Molecular biology Computer science Bisulfite sequencing Artificial Gene Amplification and Extension Bombs Biochemistry Polymerase Chain Reaction chemistry.chemical_compound 0302 clinical medicine Plasmid Nucleic Acids Community Page Medicine and Health Sciences Public and Occupational Health Biology (General) DNA extraction Materials Throughput (business) General Neuroscience Traumatic Injury Risk Factors High-Throughput Nucleotide Sequencing Amplicon Microspheres visual_art Physical Sciences visual_art.visual_art_medium Engineering and Technology RNA extraction Weapons General Agricultural and Biological Sciences Open platform QH301-705.5 Materials Science Equipment Nanotechnology Biology Bead Biomolecular isolation General Biochemistry Genetics and Molecular Biology 03 medical and health sciences Extraction techniques Coatings Animals Humans Protocol (science) General Immunology and Microbiology Surface Treatments Biology and Life Sciences RNA DNA DNA isolation Research and analysis methods genomic DNA Magnetic Fields Molecular biology techniques 030104 developmental biology Manufacturing Processes chemistry Medical Risk Factors Nucleic acid Biochemical engineering 030217 neurology & neurosurgery |
| Zdroj: | PLoS Biology, Vol 17, Iss 1, p e3000107 (2019) PLoS Biology |
| ISSN: | 1545-7885 |
| DOI: | 10.1101/414516 |
| Popis: | 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. This Community Page article presents open-source nucleic acid purification and handling protocols based on functionalised magnetic beads, with the aim of transforming life science research practice and its economics. |
| Databáze: | OpenAIRE |
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