Can anybody help me how can I amplify Bacillus cereus phage? I have 1 ml stock Of phage which was picked up after plaque assay and dropped to 1 ml Sm buffer, I have tried to pour 500 ml of stock into 5ml overnight of culture but no lysis, I wanted to make sure the phage is working and I did the plaque assay again from the stock and there was many plaques
thank you so much Peter,I did Inculate 4 ml of media with 500 ml of over night Bacillus cereus and after 1 hour I poured 500 ml of stock and waited over night and I checked it by Specterophotometer but no lysis. I will start from plaques,thank you
some times phages may loss their tails for absorption due to shaking or while petting, i am also faced the same problem, stock phage filtrate was used and after formation of plaques pour excess of SM buffer and incubate for 4-6 hrs and drain the sm buffer and added to 3-6 hr old host bacterial broth at 140-230 rpm and incubated for 18-24 hrs take OD at 600nm with and with out phages in a broth culture.
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