Rationale: To isolate possible bacteriophages through the process of soil washing and enrichment. The soil metadata will also be found and recorded.

Description of Procedures:

1. The work space was cleaned using aseptic technique and an ethanol burner was lit to create an aseptic zone.
2. 8mL of LB broth was added to the 15mL tube containing 2 mL of soil B, for a total of 10 mL. The tube was labeled LIP 9-10-18 Soil B.
3. The tube was then shaken for 10 minutes, and massed. The mass was found to be 18.212 grams.
4. While the tubes were being shaken, the some of the metadata was collected. The procedure to calculate the percent water was started. The weigh boat was found to be 2.36 g, and the weigh boat with the soil was found to be 7.27 g. The weigh boat was labeled LIP 9-10-18 Soil Dry, and was placed under the vented hood where it will sit for 48 hours to allow the water to evaporate.
5. The tube filled with LB broth and soil was put in the centrifuge for 5 minutes and spun at 10,000 x g. It was paired with a tube of similar mass (within 0.05g).
6. After the tube was spun in the centrifuge, the supernatant was filtered using a 2mL syringe and a 0.22 um filter. Approximately 7mL of lysate was collected in a 50 mL tube. There was not enough to create a direct lysate. 0.5mL of arthrobacter was added to the lysate to create the enriched lysate. The tube was labeled LIP 9-10-18 Enriched (Soil B).The tube will be shaken at 28 degrees C until Wednesday (approximately 48 hours).
7. The pH of the soil was then found. A small amount of soil was put into a pH vial and DI water was added to fill the vial. The vial was then shaken for 10 seconds and allowed to sit for 10 minutes. The pH paper was then put into the vial for 45 seconds and the paper was then compared to the pH color scale. The pH was found to be 6.0.
8. The table was then cleaned using aseptic technique and the materials were properly stored and disposed of.

Observations/Data/Results:

• Observations:
  • The filtering process was difficult and took a long time.
  • There were no more large tubes for determining percent silt, sand, and clay, so this procedure will be completed Wednesday for more metadata.
  • The cap of the pH vial had a small amount of soil stuck to the top, which could affect the results of the pH test.
• Results:
• pH: 6.0
• 7.5 mL of enriched lysate was made (0.5 of this is arthrobacter)
• The weigh boat mass: 2.36 g
• Weigh boat with soil mass: 7.27 g

Interpretations/Conclusions/Next steps:

The procedure for enrichement was completed, but there was not enough to create a direct lysate. The soil metadata was not complete and will be completed Wednesday. The percent water will also be finished Wednesday after the water evaporates for 48 hours. The next step will be to finish metadata and perform a spot test and a plaque assay with the enriched lysate.