Rationale: Due to my lack of time to both purify and amplify my phage I will conduct three plaque assays this class with 20µL, 40µL, and 80µL in order to try to get a high titer lysate sooner.

Procedure:

1. Enriched three vials of 0.5mL of arthrobacter with 20µL, 40µL, and 80µL of lysate and labeled 3 plates with corresponding values.
2. Waited for 15 minutes then begun making top agar.
3. Mixed 8mL of LB broth, 90µL of CaCl2, and 10mL of 2xTA in a 50mL tube.
4. Mixed by pipetting up and down.
5. Poured 4.5mL of top agar into control plate.
6. Aliquot 4.5mL of top agar in to each tube of arthrobacter and lysate and mixed by pipetting up and down.
7. Poured each tube into corresponding plate then swirled around to cover entire plate.
8. Waited for 15 minutes before inverting then placed into incubator.

Observations:

All plaque assays turned up negative.

Conclusion and Next Steps: I will try to run PCR on this sample, and if that also turns up negative I will continue to work on Katherine’s lysate to try to get a high titer.