Rationale:

After the flooded plate lysate goes through a filter, a plaque assay will be performed with 524 µL of lysate to make a web plate.

Procedure:

1. Filtered lysate from flooded plate “KEA 10⁰10/29” through a 0.22 µm syringe filter into a conical vial labeled “KEA 11/2/18 FS lysate 10⁰.”
2. 524 µL of “KEA 11/2/18 FS lysate 10⁰” was added to a test tube which already had 0.5 mL of Arthrobacter.
3. 4 mL of LB Broth, 45 μL of 1 M CaCl₂, and 5 mL of 2X TA were combined into a different conical vial.
4. Transferred and mixed 4.5 mL of the Top Agar mixture from the conical vial into the test tube with the Arthrobacter and enriched lysate.
5. Poured the test tube mixture onto a plate labeled “KEA 11/2 Web.”
6. The rest of the conical vial mixture was poured onto the TA control plate.
7. The “KEA 11/2 Web” plate was placed in the incubator at room temperature.

Observations:

• The following calculations were performed to determine necessary amount of LB Broth, 2X TA, and CaCl₂needed for 2 plates.

Next Steps:

If contaminated, a plaque assay will be performed again with 524 µL of the filtered 10⁰ lysate. If not contaminated, the titer will be calculated and the next steps will be determined.