Kit for antibacterial effect of T4 virus and detection by PCR amplification

Topics covered:
- Discovering bacteriophages, essential actors in the balance of microbiota
- Performing an antibiogram
- Performing a PCR amplification of a virus

Antibacterial effect of T4 virus:
Comparing in vitro the bactericidal effect of bacteriophages to that of two antibiotics. This activity follows the principle of an antibiogram. A culture plate of E. coli is divided into 4 zones: 2 antibiotics are tested in comparison to a drop of a solution (bacteriophages), the 4th quarter is the control zone.
The experiment lasts 1 session of 60 minutes, with a 3-day bacteria incubation period and 1 session to read the results. Reagents are provided for 40 plates.

PCR for detection of T4 virus:
The results of Experiment1 show that an element, not visible under the microscope and present in the deposited suspension, produces zones of inhibition of bacterial growth equivalent to some antibiotics. The PCR analysis will determine the presence of a viral gene in the suspension and thus that this bactericidal effect is due to a virus: bacteriophage T4.
The experiment lasts 2 sessions of 60 minutes. Reagents are provided for 20 PCRs.

 From the bacteriophage solutions of the two experiments, it is possible to titrate the bacteriophages in suspension with PBS buffer. This allows visualizing that each micro-lysis plaque is the result of the action of a single virus.

Composition:
- Antibacterial effect: 1L Mueller Hinton Medium, 1 tube of lyophilized Escherichia coli inoculum (k12 type), sterile water (50 ml), T4 bacteriophages, Antibiotic disks (penicillin and tetracycline), 40 Petri dishes, 40 Pasteur pipettes 1 ml, 40 spreaders
- PCR: Ready-to-use primers, PCR Mix [Nucleotides + Taq polymerase], T4 bacteriophages, DNA release, molecular weight marker, extraction buffer, 20 PCR microtubes
- Titration: PBS buffer
Additional equipment needed: P20 Micropipettes and tips, phorEasy electrophoresis system ref. 708150, Peltier module thermocycler ref. 591110, TAE buffer, agarose, GelGreen® dye