Date: Wednesday, October 10th, 2018

Title: Soil C Metadata

Rationale: The purpose of today’s lab was to record soil metadata for Soil Sample C collected 9-17-18.

Class Question: Is there a difference in bacteriophage presence or type in soil samples taken from live oaks vs those from red oaks?

Procedure:

1. An aseptic zone was set up.
2. A pinch of Soil Sample C was added to a pH vial with DI water and shaken for ten seconds.
3. The pH vial was allowed to sit for two minutes before a strip of pH paper was added to the vial.
4. After 45 seconds, the pH paper matched a pH of 6.
5. Next, Falcon tubes were filled with soil to the 10 mL mark and DI water to the 30 mL mark.
6. 3 drops of dispersion fluid were added and the tub was shaken for 30 seconds.
7. The supernatant from the Falcon tube was added to a second Falcon tube and both tubes were left under a fume hood to allow dispersion of sand, silt, and clay.
8. To calculate percent water of the sample, a weight boat was used.
   1. Empty Weigh Boat: 2.432 g
   2. Weigh Boat with Soil: 7.452 g
   3. Wet Soil: 5.020 g
9. The weight boat with wet soil was left under a fume hood to dry out.

Observations: The plaque assay from last lab yielded negative results, meaning the plaques were either incorrectly picked or they were actually air bubbles.

Results: This lab yielded soil metadata that can be used to find trends in bacteriophages.

Next Step: The next step is to wash and enrich the soil in order to get a direct and enriched sample, then to make a spot test with the enriched sample, the direct isolation, and phage buffer to test for plaques.