bioRxiv (Cold Spring Harbor Laboratory) · 2018 · 12 citations · 26 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
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 and total RNA from different sources, as well as environmental TNA and PCR amplicons. 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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Rapid and simple method for purification of nucleic acids
René Boom, C. J. A. Sol, M M Salimans et al. · Journal of Clinical Microbiology · 1990 · 5.3K citations · Full text
Central Dogma of Molecular Biology
Francis Crick · Nature · 1970 · 3.4K citations