PhiX:Plaque assay

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Revision as of 06:27, 20 February 2017 by Ben (talk | contribs) (Summary)

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Summary

This protocol is usually used in conjunction with serial dilution of a stock solution of phage for enumeration of phage titer. But it can also be used with any phage-containing input.

Media Preparation
The usual media for plating is lysogeny broth (LB). Remember that this is not a defined media (so you should attempt to minimise batch effects for reproducibility) and it comes in Lennox and Miller varieties. The appropriate type is Miller which contains 10g/L of sodium chloride (versus 5g/L in Lennox). Prepare one litre of LB:


Plating Procedure
Add 100ul

Recommended Reading
I recommend checking:
Kropinski, A.M., Mazzocco, A., Waddell, T.E., Lingohr, E. and Johnson, R.P., 2009. Enumeration of bacteriophages by double agar overlay plaque assay. Bacteriophages: Methods and Protocols, Volume 1: Isolation, Characterization, and Interactions, pp.69-76.
for more details of the plaque assay and variations thereon.