I have 17 phages from sewage and want to test them on 50 bacterial strains obtained from UTI (urinary tract infected) patients as a means of characterization based on host range, i also want to test the phages sensitivity to chloroform, different pH, and temperatures. I dont know exactly how to approach this characterization! Ány ideas would be helpful, thank you everyone
Depending on which instruments you have available you could use a microplate reader, grow cells and add phages, reading the absorption at 600 nm. Upon lysis the absorption will drop dramatically. The same could be done manually (record OD at 600 nm) but would be way too much with your number of phage / bacteria combinations. Another way could be a luminescence assay (luciferase) detecting for ATP. Also available in a microplate reader format + luminometer. Upon cell lysis there is still ATP but the amount should decrease significantly over time.
Having established the host range, you have to repeat the positive hits with phage lysate that has been treated with the parameters you mentioned: chloroform, pH extremes and temperatures.
Hope that helps - good luck!
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