Hi, I'm working on a project where we're using S. aureus and corresponding bacteriophages. I did a spot plate and found out that my phage stock is roughly 2.0x10^7 PFU/ml. I want it to be at least 10^10 PFU/ml. My method for growing the bacteriophages has been to make agar plates and then use top agar that's been mixed with the bacteria and the phage dilution. How will I go about making more stock solution that is the appropriate concentration? Do I just have to make it and then test it?
Thank you.
to get the highest titer from a plate stock, you have to have the phage/cell ratio optimal. I would make a series of plates with different dilutions of your phage stock and constant amounts of cells, prepare one plate from each, and collect the phage from each. Then you can do a quick titer from each stock and save the one or two that are the highest.
Tangential flow filtration works well for phage concentration. https://www.researchgate.net/publication/281304462_Evaluation_of_Tangential_Flow_Filtration_for_the_Concentration_and_Separation_of_Bacteria_and_Viruses_in_Contrasting_Marine_Environments
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