PLoS Biology · 2019 · 320 citations · 30 references
Life SciencesBiotechnologyMolecular BiologyBioseparationsMolecular BiotechnologyNucleic Acid BiochemistryMagnetic SeparationNucleic Acid EngineeringNucleic Acid ExtractionNucleic Acid PurificationBead-based AssaysBead-based SynthesisGenomic Dna AnalysisGenomic Dna PurificationHigh-throughput ExtractionHigh-throughput PurificationMagnetic Bead PurificationMagnetic Bead-based Extraction
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.
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Molecular cloning: A laboratory manual
Prescott L. Deininger · Analytical Biochemistry · 1990 · 86.2K citations