When I was amplifying the phage library, TG1 cells with an OD600 of 0.4-0.6 were infected with M13KO7 at an MOI of 20 and incubated at 37°C for 1 hour. Subsequently, I centrifuged the cells and resuspended them in medium containing Amp (the resistance marker carried by the phagemid) and Kan, then cultured them at 25°C. However, after 3 hours of cultivation, we initially observed a significant decrease in the OD of the bacteria, and after overnight culture, many bacterial fragments were found in the medium. I would like to know what might be causing this issue. Thank you for your response, and best wishes!
It is hard to guess from your experiment but either you have a different lytic phage contaminating something, or you may be losing your phagemid and the cells are lysing due to the amp. Is the phagemid strain growing well with amp in the absence of M13K07?
The death of the bacteria shows that the phage has affinity for the bacteria and is capable of inducing lysis.
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