Rationale: Collect a new soil sample, start collecting the Soil Metadata for the soil sample, and create a enriched lysate for a plaque assay. 

Procedure: 

1. Created an aseptic zone using an ethanol burner after cleaning the table with Cidecon and 70 % ethanol.  
2. Collected 2mL of soil in a 15 mL vial and filled another 50 mL vial with soil.  
3. Added 10 mL of LB broth into the 2 mL soil vial, and shook for 10 minutes  
4. In the meantime, obtained soil metadata measurements for percent water. 
5. Set up the soil metadata for percent sand, silt.  
6. Obtained a falcon tube and added 10mL of soil 
7. Added DI water till the 30mL mark.  
8. Added a few drops of soil dispersion liquid.  
9. Shook for a minute and set it in the hood to be observed next time  
10. After 10 minutes of shaking the LB Broth and soil, centrifuged for 10 minutes at 5,000xg. 
11. Collected PH of soil by filling tube with little soil and water and let it sit for 10 minutes.  
12. After a few minutes placed the ph paper for 45 seconds and observed the results.  
13. Obtained the soil and LB broth solution from the centrifuge. 
14. Filtered the supernatant into a 50 mL vial using a syringe and ended up obtaining about 7 mL of enriched lysate.  
15. Added 0.5 mL of Arthro into the enriched lysate.  
16. Placed it in the shaking incubator until next class.  

Observations/ Results: 

Plaque Assay from 10/08/18 

• 3/5 of our plates were contaminated with a red pigmented bacteria, and control was also contaminated
  

  Plate results were negative : no contamination or plaques

  

  Control Plate was contaminated

  

  Soil Sample C was contaminated with red pigmented bacteria

  Soil Metadata for Soil E: 

  % water : 

  • Weigh boat = 2.475 grams 
  • Weight boat + soil = 5.817 grams 

  PH of soil = 6.0 

Next Steps:  

We will be performing a plaque assay using the new soil sample: Soil Sample E. We will also collect the remaining soil metadata for this soil sample.