Isolation of Bacteriophages Against Drug-Resistant Bacteria

Begin with a sewage water sample containing bacteriophages.

Add growth medium and drug-resistant bacteria, then incubate.

Host-compatible bacteriophages attach to host cells and inject their genetic material, while incompatible phages fail to establish infection.

Inside the cytoplasm, the genetic material replicates and directs the production of viral proteins.

Bacteriophage particles assemble in the cytoplasm, and the host cell then lyses, releasing them into the surrounding medium.

Centrifuge to pellet the debris.

Transfer the bacteriophage-rich supernatant to a new tube.

Add fresh medium and drug-resistant bacteria, then incubate to increase bacteriophage production.

Centrifuge again to pellet the debris.

Mix the bacteriophage-containing supernatant with drug-resistant bacteria in molten soft agar, overlay it on an agar plate, and incubate.

Observe clear zones, called plaques, on the bacterial lawn, indicating bacteriophage-mediated bacterial lysis.

Pick an individual plaque and transfer it into growth medium for repeated plaque purification to isolate a homogeneous bacteriophage population targeting the drug-resistant bacteria.