Collecting Metadata and Plaque Assay 10/3/18

Rationale: Conducting a plaque assay in hope of getting a plaque. Collecting metadata in order to eliminate some confounding variables.

Procedure:

1. Spun lysate at 3000g for 5 minutes then filtered 1.5mL with 22nm filter andsyringe.
2. Added 10μL of lysate to 0.5mL arthrobacter.
3. Setup soil composition analysis by adding 10mL of soil to falcon tube thenfilling to 30mL with DI water. Added 3 drops of soil dispersion fluid and letsit.
4. Put 3.24g of soil in a weight boat of 2.40g and let dry under vent hood.
5. Conducted soil pH test.
6. Made top agar with 4mL LB broth, 45μL CaCl2, and 5mL of 2x TA.
7. Poured 4.5mL into a control plate then poured arthro and lysate solution intoremaining top agar.
8. Poured the top agar into a plate and let sit. Test plate slid around and didn’tfully solidify, but control plate did not. Put test plate in incubator right side up and control plate reversed.

Observations:
pH of soil was between 5.5 and 6.0. Soil contained 67% silt and 33% clay. The amount of sand was too small to be measured quantitatively. Plaque assay didn’t solidify properly due to not letting plate get to room temperature before pouring.

Interpretations and Next Steps: This tree has had the most acidic soil I have seen thus far. I need to run another plaque assay no matter what next lab b because whatever results I get will be hard to read.