Abstract
Viruses influence ecosystem dynamics by modulating microbial host population dynamics, evolutionary trajectories and metabolic outputs. While they are ecologically important across diverse ecosystems, viruses are challenging to study due to minimal biomass often obtained when sampling natural communities. Here we describe a technique using chemical flocculation, filtration and resuspension to recover bacteriophages from seawater and other natural waters. The method uses iron to precipitate viruses which are recovered by filtration onto large-pore size membranes and then resuspended using a buffer containing magnesium and a reductant (ascorbic acid or oxalic acid) at slightly acid pH (6–6.5). The recovery of bacteriophages using iron flocculation is efficient (>90%), inexpensive and reliable, resulting in preparations that are amenable to downstream analysis by next generation DNA sequencing, proteomics and, in some cases, can be used to study virus–host interactions.
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Acknowledgments
We thank Jennifer Brum and numerous viral ecologists for their feedback about this evolving protocol and its application across diverse sample types. This work was supported by funds from the Gordon and Betty Moore Foundation through a collaborative (GBMF3305) to M.B.S. and S.G.J., as well as methods development grants (GBMF2631 and 3790) to M.B.S.
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Poulos, B.T., John, S.G., Sullivan, M.B. (2018). Iron Chloride Flocculation of Bacteriophages from Seawater. In: Clokie, M., Kropinski, A., Lavigne, R. (eds) Bacteriophages. Methods in Molecular Biology, vol 1681. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7343-9_4
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DOI: https://doi.org/10.1007/978-1-4939-7343-9_4
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