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How can a mixture of unspecified phages be separated (into singular phage strains)? I.e, what are the main methods?

My research shows it can be done using CsCl centrifugation and affinity chromatography, but both seem laborious, the latter also requiring the phages to be already available in pure form.

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Look up "phage purification." Basically, you add your phage solution on top of your host bacteria on a plate and incubate. As the bacteria grow, you should see plaques (clear areas in the bacterial lawn). Each plaque should be the progeny of one original phage, although sometimes two or more phages could be very close to each other on the plate and it will look like one plaque. So you have to take a sample of a plaque, re-plate on another lawn of host bacterium, and repeat this process for several rounds until the plaque morphology seems to indicate that there is only one type of phage present. Here is a good resource for phage protocols: http://phagesdb.org/workflow/

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  • $\begingroup$ Thanks, excellent link! Any other known way other than manual extraction from plate? Is it perhaps easier to isolate infected bacteria during lytic cycle (or perhaps during induced lysogenic cycle)? $\endgroup$ – Zubo Sep 21 '17 at 0:31

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