Rationale: I am conducting a plaque assay to confirm whether or not the plaque I found in my spot test done on soil sample B from tree A was truly due to the presence of a bacteriophage.

Procedure:

1. Mixed diluted enriched lysate made in the previous procedure with 0.5mL of arthrobacter

   and let sit. Totaled about 12 minutes of letting sit before being poured in the plate.

2. Used aseptic technique to create an aseptic zone.
3. Made top agar by mixing 8mL of LB broth, 90μL of CaCl2 and 10mL of 2X top agar.
4. I pipetted 4.5mL of the solution into my tube of arthrobacter and enriched lysate and

   pipetted up and down to mix.

5. I then poured the mixture onto the plate and let dry for 10 minutes.
6. Due to a lack of plates we split a plate in four and poured 1mL of our top agar without

   any arthrobacter into one of the quadrants to use as a TA control.

7. I stored my plate in the incubator for 48 hours.

Observations:
Plaque assay returned negative.

Next steps:
Due to the negative results of the plaque assay I will have to go look for more soil and test once again.