Rationale: Set PCR to amplify the phage DNA to prepare for DNA GEL. DNA GEL will determine the presence of phage in the obtained soil sample A.

Procedure: Before the experiment was performed, the workspace was cleaned with both Cidecon and 70% Ethanol. A burner was then placed to set up the aseptic zone. The lysate was obtained and 1mL was added to a microcentrifuge cap. This boiled for 10 minutes to break up the protein ca[sid so the phage DNA could be extracted. In three microcentrifuge caps, Primers 1,2, and 3 were added. The formula below shows the solution concentration added to the caps (x3 for 3 microcentrifuge caps):

• 12.5 microns Taq
• 2 microns of phage DNA
• 6.6 microns of DH2O

After the solutions were added, the three caps were placed in a thermocycler for 52.5 minutes.

Observations: Control and the experiment performed on 10/22 were both negative. No plaques were present on the plaque assay performed on 10/22. No plaques were available to pick, and PCR was done to test whether the soil sample is positive or negative.

Negative Control 10/22

Negative experiment 10/22

Conclusions: Lab experiment performed on 10/22 was negative due to contaminations. Possible contaminations reasons:

• LB broth and 2X TA are contaminated
• Opening/Closing of microcentrifuge caps/things not done in an aseptic zone.

The lysate was used to perform PCR in preparation for DNA GEL experiment. DNA GEL will test whether or not the soil sample obtained is positive/negative. If the soil sample is negative, new soil will be needed to be tested via PCR/DNA GEL.