Rationale:

To conduct a PCR test in order to determine if Soil F could potentially contain a phage. Additionally, finish collecting metadata on Soil F in order to understand more about the soil type.

Procedures:

1. Setup an aseptic zone.
2. Centrifuged Soil F Enriched Lysate and transferred 1mL of the supernatant to a micro test tube, “NMN 10.22 EL-F PCR”.
3. Analyzed the falcon tube for soil composition.
4. Analyzed the percent water test.
5. Boiled “NMN 10.22 EL-F PCR”.
6. Added 6.5µL DDI water, 1µL EL-F PCR, 1µL of Rams Enriched Lysate, 4µL Primer 1, and 12.5µL of Taq Polymerase to a microtube. Labeled with a circle and number to indicate Primer type.
7. Repeated step 6 for Primers 2 and 3.
8. Added 7.5µL DDI water, 1µL Control DNA, 4µL Primer 1, and 12.5µL of Taq Polymerase to a microtube. Labeled with a plus sign and number to indicate Primer type.
9. Repeated step 8 for Primers 2 and 3.
10. 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. I also observed 6mL Sand, 1.5mL Clay, and 2.5mL Silt in the falcon tube. Thus Soil F is 60% Sand, 15% Clay, and 25% Silt, making it sandy loam. I also recorded that the dried weight of only Soil F was 3.353g, thus Soil F is 37.09% water.

Conclusions/Next-Steps:

From the data collected, I can conclude that Soil F was fairly moist at roughly 37% water and it was of a sandy loam variety. THe next step will be to conduct a gel electrophoresis test in order to confirm or deny the presence of phage in my Soil F and/or Ram’s soil sample.