Rationale: To conduct a PCR to determine whether or not there are phages and to amplify the DNA of the phages if they are present within the enriched lysate

Procedure:

• To prevent contamination, wipe the table with CiDecon and ethanol and set up an aseptic zone.
• Due to the presence of no phages, another method was used to determine if there were phages in the sample was taken, PCR.
• First, the sample was boiled.
• 1 µL of the enriched lysate ( in each of the three tubes) was added to the 2X Master Mix (12.5 µL of Taq polymerase and dNTP)
• 4 µL of primer was added to their respective tubes.
• 6.5 µL of ddH2O was added to each tube.
• For the control, 7.5 µL of ddH2O and 4 µL of primer 3 were combined.

Results and Analysis:

On 10^-2, the clear spaces are due to the top agar being torn

Conclusion:

Because there were no phages on the plates, another approach to check for phages was taken. The first step to conduct PCR was to boil the enriched lysate was 95 degrees Fahrenheit. Then, 1 µL of the enriched lysate was added to the 2X Master Mix along with 6.5 µL of ddH2O. Primer was added to their respective tube. This was conducted three times, adding each component in equal volumes.

Future Plans:

If there are phages present in the sample, the DNA will be amplified and will be used for a plaque assay to pick a plaque.