Rationale:

To conduct a gel electrophoresis test in order to test the effectiveness of the PCR procedures, by testing a PCR sample with a known positive for the phage and seeing if the results of the gel show a positive presence for phage DNA.

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. Added 5µL of ladder to a well in the apparatus.
6. Added 2.5µL of 10x DNA gel dye to all PCR tubes from Monday.
7. Pipetted 5µL of all PCR tubes into different wells.
8. Attached a power source and ran the test for 40 minutes.
9. Analyzed the gel in the gel imager.

Observations/Data:

The experimental gel came back negative and the control gel performed as expected. The ladder also performed as expected.

Conclusions/Next-Steps:

Based upon the data collected it can be concluded that the PCR procedure is faulty as the gel came back negative for phage DNA when it was known that there was phage DNA in the sample used. The next steps will be to help Claire in producing a high titer lysate that can be imaged with TEM and ultimately sequenced.