The lab I work in is looking to isolate phages from water sources, but we have had no luck in the past. What is the best protocol for isolating phages from water samples? We work primarily with Mycobacterium hosts.
Hi James Bruns .
Filter the water using filter paper to separate coarse materials. Then, consider doing serial dilutions to isolate bacteria on LB agar plate and purify the bacteria to obtain pure colonies.
To isolate phages, try spot test method. Mix bacteria culture together with the water sample and LB, centrifuge and filter using 0.22 μm. Next, do the standard plating by transfer 100 μL of the filtrate and 100 μL of the bacterial culture onto 1/2 LB agar plate, pour 3 mL of Top agar on top of it (you can adjust the quantity where applicable) and incubate overnight. Observe the clear zones of phages on the next day. All the best!
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