Rationale: After two using soil C to make three enriched lysates and only finding plaques from one of them, the purpose of this lab is to collect new soil from the same tree as soil C and enrich this soil for future testing.

Description of Procedures:
Soil C was recollected from the same tree. 2 ml of soil and 10 ml of LB broth were added to a tube and shaken for 10 minutes. While it was being shaken, the percent water calculation for the soil was begun. The weigh boat was found to be 2.33 g. With the added soil it was found to be 6.388 g. The weigh boat was then stored under the vent hood and will remain there to allow water to evaporate until the next lab. Next, the test for the composition of the soil was started. 10 ml of soil and 20 ml of DI water were added to a tube with 3 drops of soil dispersion liquid. The tube was shaken for 30 seconds and then placed under the vent hood until the next lab. After shaking the tube, the tube was spun in the centrifuge at 5000 x g for 10 minutes. While spinning, the pH of the soil was found. Soil and DI water were added to a pH vial and allowed to sit for 10 minutes. When complete, the pH was found to be 5.5. After spinning in the centrifuge, the supernatant of the soil collection was filtered with a 0.22 um syringe filter. Approximately 7.5 ml of lysate were obtained. 0.5 ml of arthrobacter were added to this tube for enrichment. The tube was labeled LIP SS EAG Soil C Repeat 10-3-18. The tube will be shaken at room temperature until the next lab. The workstation was cleaned using aseptic technique and the materials were properly stored or disposed of.

Observations/Results:

• Observations:
  • Soil C repeat floated before being shaken.
  • Soil C repeat had a lower pH than the original Soil C.
• Results:
  • pH: 5.5
  • 7.5 ml of lysate obtained

Interpretations/Next Steps:

The procedure was complete. The next step will be to finish the soil metadata for Soil C repeat and to run a plaque assay with Soil C repeat.