I have a trouble isolating bacteriophage from the anaerobic bacterial host of Clostridium perfringens. First of all, I have already use various method including doble layer agar method and Spot test method with various concentration of agar concentration and type of media used. As far as i concern, various paper suggest the usage of Brain-Heart Infusion (BHI) as medium but I still can't archive a proper bacterial lawn, not to mention a single clear phage plaque. Can someone enlighten me?
Are you maintaining anoxic conditions during growth (reducing agents in media, incubation with gas packs)?
Jakub Grzesiak I did maintain the anaerobic condition of the host as well as maintain the optimised condition for the host. I have successfully isolated the phage recently and going to do the subsequent characterization. Thank you so much for the insight.