Rationale: Perform a plaque assay and collect the remainder of soil metadata: percent water, percent sand silt and clay.  

Procedure: 

1. Cleaned table with Cidecon and 70% ethanol and created an aseptic zone with an ethanol burner. 
2. Obtained weigh boat with soil from hood and obtained percent water results.  
3. Obtained the enriched lysate, and centrifuged for 10 minutes at 5,000x g. 
4. While lysate was being centrifuged, collected tube from hood to collect percent sand, silt and clay for soil.  
5. Collected enriched lysate from centrifuge and filtered with syringe into a 50 mL vial. 
6. Resulted in about 6 mL of enriched lysate. 
7. Added 112.5 Cacl2, 10 mL of LB Broth to another 50 mL vial.  
8. Added 10 uL of enriched lysate to 0.5 mL of Arthro vial.  
9. Let it sit for 10 minutes.  
10. Added 12 mL of 2x TA into the solution in the 50 mL vial.  
11. Added 4.5 mL of solution into each of our plates.  
12. Let the plates sit for 10 minutes.  
13. Placed the plates inverted in the incubator.  

Observations/ Results  

Soil Metadata 

• Percent water for Soil D = 11.48% 
• Sand =  (6.25mL / 8mL) = 78.13% 
• Silt =  (0.75mL / 8 mL) = 9.37% 
• Clay =  (1ml / 8mL) = 12.5%
  

  Soil D Metadata

While plating today there were bubbles in the control plate.  

Next Steps: 

Next time we will observe the results of the plaque assay plates. If the plates are negative then we will collect a new soil sample, enrich the soil sample, and start collecting soil metadata. If the plates are positive, we will continue with amplifying and purifying the plaque.