Rationale

PCR will be conducted on an enriched isolation of soil sample C produced on 10/24/18.

Procedure

• Established an aseptic zone.
• The enriched isolation produced on 10/24/18 was centrifuged at 5000g for 5 minutes.
• 1 mL of the spun enriched isolated was aliquotted into a microcentrifuge tube and was heated for 10 minutes.
• 4 µL of primer mix 1, three 2 µL samples of “phage DNA”, 2.5 µL of DDI water, and 12.5 µL of TAQ polymerase were added to PCR tube 1.
• 4 µL of primer mix 2, three 2 µL samples of “phage DNA”, 2.5 µL of DDI water, and 12.5 µL of TAQ polymerase were added to PCR tube 2.
• 4 µL of primer mix 3, three 2 µL samples of “phage DNA”, 2.5 µL of DDI water, and 12.5 µL of TAQ polymerase were added to PCR tube 3.
• The 3 tubes were then cycled through PCR and stored for later use.

Observations

The mixture in the PCR tubes was colorless rather than the teal color that was observed at the last round of PCR. This is due to a lack of dye that will be added later before conducting gel electrophoresis.

Conclusions and Next Steps

Using the PCR results, gel electrophoresis will be run on Monday to test for phage presence in the sample.