Can somebody share a standardised protocol for isolating phages against gram-positive bacteria
Hi Moein,
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, centrifuge it, filter it, and do a plaque assay with the same host. Keep the above suspension back at 30 C overnight.
3) After overnight, again do centrifugation, filtration and plaque assay.
Using this protocol, you will get phages with distinct plaque morphology without any phage with a very low population.
In case you don't get the phages, use a different sample and follow the same protocol.
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