JoVE Encyclopedia of Experiments
Microbiology
0 views • 4:30 min • July 31st, 2026
Take a suspension of Borrelia burgdorferi bacteria infected with a borrelial phage.
Centrifuge the suspension and collect the supernatant containing phage particles and cellular debris.
Add a salt solution and incubate with shaking.
The salt solution enables the separation of residual host-derived macromolecules from the phage particles.
Centrifuge the suspension and collect the supernatant containing the phage particles.
Add polyethylene glycol or PEG, which acts as a molecular crowding agent that facilitates phage particle precipitation.
Incubate on ice and then centrifuge.
Discard the supernatant.
Resuspend the pellet in a buffer, then transfer it to a microcentrifuge tube.
Treat with chloroform to dissolve lipid-based cell debris.
Centrifuge the tube and collect the upper aqueous phase containing the borrelial phages.
Repeat the chloroform addition, mixing, centrifugation, and collection steps to ensure complete removal of contaminants.
Store the isolated phage particles under cold conditions for further use.
Supplement the prepared culture with the appropriate antibiotic concentration described in the manuscript, while incubating the sample at 33 degrees Celsius for 72 to 96 hours. To prep
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