Objective:

• Wash the soil sample (Soil A) to remove unnecessary material, such as dirt, rocks, and bacteria.
• Isolate possible bacteriophage found in the soil, in lysate.

Procedure:

1. Aseptic Zone was prepared. Lab space was cleaned using CiDeon and wiped dry with paper towel. Ethanol (70%) was then sprayed, wiped, and evaporated. Ethanol burner was then lit on the table.
2. 50 mL tube was obtained that contained the sample of Soil A up to the 12 mL mark. LB broth was then poured into tube up to 35 mL mark.
3. The mixture in the tube was shaken manually for 15 minutes.
4. After mixing, the tube was massed in order to find another tube with a similar mass (+/- 0.1g) that it could be placed with in a centrifuge. The 50 mL tube was found to be 55.215 g.
5. 50 mL tube containing mixture of Soil A and LB broth was centrifuged for 5 minutes.
6. Liquid found at the top of the soil in the tube was then filtered using a top filter and vacuum. This was to remove the bacteria from the remaining soil. 10 mL of filtrate was obtained from the filter in a 50 mL tube.
7. Half (5 mL) of the filtrate was put into a 15 mL conical vial and placed in the fridge. This was the direct lysate. Vial was labeled Direct Soil A.
8. 0.5 mL of Arthrobacter was added to the remaining lysate (5 mL) in the 50 mL tube. This created the enriched lysate. 
9. Enriched lysate was left to be shaken to be shaken until Monday (8/27).

Observations:

• After the soil sample was centrifuged the liquid was still murky. This may have lead to difficulty during the filtering,
• Filtering the lysate was difficult and took longer than other samples. Due to time, only 10 mL was obtained instead of all lysate that was available.

Results:

• No results to report since test has not been completed.

Next Steps:

• In the next lab, spot tests and plaque assays will be completed to see if bacteriophages are present in the soil sample, Soil A.