Bio-On-Magnetic-Beads (BOMB): Open platform for high-throughput nucleic acid extraction and manipulation.
Phil OberackerPeter StepperDonna M BondSven HoehnJule FockenVivien MeyerLuca SchelleVictoria J SugrueGert-Jan JeunenTim MoserSteven R HoreFerdinand von MeyennKatharina HippTimothy Alexander HoreTomasz P JurkowskiPublished in: PLoS biology (2019)
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.
Keyphrases
- nucleic acid
- high throughput
- molecularly imprinted
- single cell
- randomized controlled trial
- healthcare
- escherichia coli
- solid phase extraction
- mental health
- drinking water
- social media
- climate change
- minimally invasive
- single molecule
- gene expression
- dna methylation
- liquid chromatography
- klebsiella pneumoniae
- high resolution
- cell free
- multidrug resistant