Rationale:

To conduct a PCR test in order to test the validity of the PCR procedure used by utilizing lysates that have been confirmed positive.

Procedures:

1. Setup an aseptic zone.
2. Transferred 1mL each of Claire’s ELF-B and Lucy’s ELF-C to micro test tubes, “Claire” and “Lucy” respectively.
3. Boiled both “Claire” and “Lucy”.
4. Added 4.5µL DDI water, 2µL “Claire”, 2µL of “Lucy”, 4µL Primer 1, and 12.5µL of Taq Polymerase to a PCR tube. Labeled with a star and number to indicate Primer type.
5. Repeated step 6 for Primers 2 and 3.
6. Thermocycled all tubes.

Observations/Data:

I observed that some of the additions to the microtubes did not immediately make contact and mix with the main body of fluid.

Conclusions/Next-Steps:

The next step will be to conduct a gel electrophoresis test in order to confirm or deny the validity of the PCR procedure by seeing if the tests come back positive as they are expected to be.