Rationale)

Using a lysate produced from Soil D I will conduct a plaque assay and a spot test in order to test for the presence of phage in Soil D. I will also collect metadata from Soil D.

Procedures)

An aseptic zone was established and the falcon tube from 9.26.18 was analyzed for soil composition, the percent H2O sample from 9.26.18 was also analyzed. Soil D Enriched Lysate was filtered with a .22µ syringe filter, 10µL of the filtrate was added to a vial of .5mL of arthro to infect for 15 minutes. The infected arthro was added to a vial containing 2mL of LB broth, 22.5µL of 1M CaCl2, and 2.5mL of 2xTop Agar, likewise 2.5mL of LB broth, 22.5µL of 1M CaCl2, and 2.5mL of 2xTop Agar was added to another vial to act as a top agar control. Both vials were plated with the prior as the plaque assay and the latter as a top agar control, the plates were incubated after solidification. Added .5mL of arthro, 2mL of LB broth, 22.5µL of 1M CaCl2, and 2.5mL of 2xTop Agar to a vial for a spot test, likewise 2.5mL of LB broth, 22.5µL of 1M CaCl2, and 2.5mL of 2xTop Agar was added to another vial to act as a top agar control. Both vials were plated into their respective plates. Spotted 10µL of Phage Buffer, Filtered Enriched Lysate, and Direct Lysate onto respective sections of spot test plate, incubated plates after solidification.

Observations/Data)

Sand: 2mL, 66%; Silt: .5mL, 16%; Clay: .5mL, 16%. Mass of weigh tray and dried soil: 5.359g; Mass of dried soil: 3.028; Mass of Water: .972g; Percent Water: 24.3%. There were some bubbles in my plaque assay, which were subsequently marked with a sharpie to prevent potential misidentification. The spot test

Conclusions/Next Steps)

From the data I collected it can be determined that Soil D can be classified as Sandy Loam and was relatively hydrated with 24.3% water. My next step will be to check for positive or negative results in both the spot test and the plaque assay, if positive the plaques will be picked and purification will begin, if negative another soil sample will be collected.