Difference between revisions of "PhiX:Plaque assay"
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==Summary== | ==Summary== | ||
This protocol is usually used in conjunction with [[PhiX:Serial dilution | serial dilution]] of a stock solution of phage for enumeration of phage titer. But it can also be used with any phage-containing input. | This protocol is usually used in conjunction with [[PhiX:Serial dilution | 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'''<br> | ||
| + | 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'''<br> | '''Plating Procedure'''<br> | ||
Add 100ul <br> | Add 100ul <br> | ||
| − | ''' | + | '''Recommended Reading'''<br> |
| − | + | I recommend checking:<br> | |
| + | 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.<br> | ||
| + | for more details of the plaque assay and variations thereon. | ||
Revision as of 06:27, 20 February 2017
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.
