i isolated the phage and put in SM buffer and than in 3ml brucela broth add 100ul bacteria and 100ul phage from SM buffer incubated for 48 hr at 42 hr than centrifuge at 10000rpm for 10 min than filtered and than i make 10 folds dilutions.i can see plaques in 3rd or 4th dilutions but in original and first dilutions. And plaques are very big i am very confused either it is phage or not and why not in higher dilution and one more thing after 42 hours incubation my double agar plate has some liquid floating i am using lower agar 1.2% with NZCYM broth and top agar 0.4% please see the images also
The problem is that you don't have good bacterial lawns so it is hard to see whether or not you have plaques. I am not sure if the large "Plaques" are really plaques or just water spots, but they look very large.
I would streak your host strain for single colonies (do it twice) to be sure you get a clean stock that isn't contaminated. Then start your host culture from a clean single colony and try again.
sir thanks for replying.Today i placed some plates straight in the incubator for 2 hours than placed inverted i have good results in that but the plates which i placed inverted from the start they all have water top agar i am using is 0.5%
These plates and plaques look much better. It does not look like the water or condensation is affecting anything, but if you are worried then be sure the plates are sufficiently dry before using. You could leave them for a day at room temp or a few hours in the incubator before you use them.
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