Rationale: Wash newly collected soil in order to get both a direct and enriched sample isolation. In addition, calculate soil metadata.

Process:

1. Wash table and set up aseptic zone
2. Added ~10 mL LB broth to ~2 mL soil
   1. accidentally poured to much and used a bulb pipette to remove ~2 mL LB broth
3. Shook and vortexed for 15 minutes
4. Mass of tube = 20.33 g
5. Centrifuged for 10 minutes
6. Filtered with syringe filter
   1. used ethanol to constantly clean tip
   2. ended with ~7 mL for enriched isolation and ~1 mL for direct isolation
7. Added 0.5 mL arthro to enriched isolation
8. Incubated enriched for 48 hours
9. Stored direct in fridge

Soil Metadata

• % H2O
  • mass of dry soil and weigh boat = 6.66 g
  • mass of wet soil and weigh boat = 6.77 g
  • mass of H2O = 0.11 g
  • mass of wet soil = 4.45 g
  • % H2O = 0.11 g / 4.45 g * 100% = 5.27%
• pH
  • added pinch of dirt to pH tube
  • filled with DI
  • shook for 10 seconds
  • held pH paper in for 45 seconds
  • pH = 5.5
• sand/silt/clay
  • total ≈ 4 mL
  • sand ≈ 2 mL
  • silt ≈ 1 mL
  • clay ≈ 1 mL
  • % sand = 2 mL / 4 mL * 100% = 50%
  • % silt = 1 mL / 4 mL * 100% = 25%
  • % clay = 1 mL / 4 mL * 100% = 25%

sand/silt/clay dispersion in falcon tube

Next steps: Perform spot test to check for any plaques.