Rationale:

To conduct a gel electrophoresis test in order to test for the presence of phage in Soil F, by either confirming that phage is absolutely not present or could potentially be present.

Procedures:

1. Added 35mL TBE, 1.7µL EtBr, and .7g Agarose to an Erlenmeyer flask.
2. The flask was microwaved to create a solution.
3. The gel solution was poured into trays and left to solidify, after solidification the dams and comb were removed.
4. The gel apparatus was filled to about 1cm above the actual gel with TBE.
5. Repeated steps 1-4 to create another apparatus
6. Added 5µL of ladder to a well in each apparatus.
7. Added 2.5µL of 10x DNA gel dye to all PCR tubes from Monday.
8. Pipetted 5µL of all PCR tubes into different wells utilizing both apparatuses.
9. Attached a power source and ran the test for 45 minutes.
10. Analyzed the gel in the gel imager.

Observations/Data:

When pipetting Primer 2 Positive Control into a well the tip went a bit deeper than it should have. The experimental gel came back negative and the control gel performed as expected, however, the lines were relatively faint. The ladder also performed as expected.

Conclusions/Next-Steps:

From the data collected it can be concluded that Soil F is negative for phage, thus the next step will be to collect another soil sample and analyze it via PCR for the presence or absence of phage DNA.