Recently, The isolation of bacteriophages from sewage water is so difficult, How to obtain phages plaques when i make isolation bacteriophage experiments for these bacterial types as bellow;
1- Psychrobacter alimentarius.
2- Bacillus tropicus.
4- Leuconostoc mesenteroides.
and which better for use, (LB), or (MRS) Media?
Hi Magdy,
Isolating some phages, especially for the gram-positive hosts, is very difficult. You may use the following protocol:
1) 5 ml of unfiltered sewage + 5 ml 2X LB Broth + 200 ul overnight grown culture in a 50 ml tube.
2) Incubate the suspension at 30 C for 6 hrs. Take 1 ml of this suspension and centrifuge, filter and do a plaque assay with the same host. Keep the above suspension back at 30 C overnight
3) After overnight, again do centrfugation, filtration and plaque assay.
Using this protocol, you will get phages with distinct plaque morphology without any phage with a very low population.
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