October 19

Soil Washing 10/15/2018

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

Process:

  1. Wash table and set up aseptic zone
  2. Added ~10 mL LB broth to ~2 mL soil
  3. Shook and vortexed for 15 minutes
  4. Centrifuged for 10 minutes
  5. 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
  6. Added 0.5 mL arthro to enriched isolation
  7. Incubated enriched for 48 hours
  8. 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 = 6.0
  • sand/silt/clay
    • added to falcon tube:
      • 4 mL soil
      • 8 mL  deionized water
      • 3 drops dispersion oil
    • shook for 45 seconds
    • let sit under hood for 48 hours

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


Posted October 19, 2018 by rachel_melone1 in category Rachel Melone

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