Hiii. I have phage 5' for E.coli ATCC 13706. I tried to increase phage concentration by method: 50 ml phage 108 PFU/ml + 10 ml of E.coli + 150 ml of PYCa medium, after incubating at 37 oC for 12 h, little clear medium can be seen, and the concentration of phage is around 108 PFU/ml.
Do you know any method to increase phage concentration to 1011 or 1012 PFU/ml? please let me know. Thank you very much.
Hi,
I recently had a lot of success with bacteriophage purification methods. The one that worked best for me is the Cesium Chloride gradient. It consists of a normal bacteriophage amplification which is then centrifuged at very high speed on a CsCl gradient. Bacteriophages are located in a thin visible band that can than be sampled. With MS2, I had titles up to 10^14 PFU/ml.
I would be glad to share my protocol with you if you are interested.
Hi Son
I scanned your method and I should work fine to increase your phage PFU, just I am concerned about the phage amount that you are adding (50 ml of 108 ) is too much so either that high number will not result in more infection and the final phage particles remain the same (try 1-10 ml) or your E-coli strain that you are using are resistant to further infection so try to use new fresh strain. Furthermore, try to remove your infection mixture after 3-4 hours if clearance of the broth is visible. Hope that will help.
Hi Clara. I did the first protocol that you suggested, it was not success.
Hi Mohammed. I will follow your idea. I hope it will be ok. Thanks.
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