Rationale: The purpose of this lab is to calculate the titer of the lysate, flood plates to collect more lysate, and create plaque assays for future flooding.

Description of Procedures: 

1. The workstation was cleaned using aseptic technique and an ethanol burner was lit.
2. The titer was calculated to be 5.1 x 10^7, a medium titer.
3. The 10^-1 and 10^-2 plates were flooded with 8 ml of phage buffer each and placed on the shaker for 1 hour.
4. 10 ul of 10^-2 lysate was added to 4 tubes of 0.5 ml of arthrobacter and allowed to sit for 10 minutes.
5. Top agar was made for 5 plates. 10 ml of LB Broth and 112.5 ul of CaCl2 were added to a tube.
6. 12.5 ml of 2x TA was added to the tube. 4.5 ml of the top agar solution was added to each of the arthrobacter tubes and poured directly onto plates labeled LIP 11-14-18 PA1, LIP 11-14-18 PA2, LIP 11-14-18 PA3, and LIP 11-14-18 PA4. The rest of the solution was poured onto a plate labeled LIP 11-14-18 control.
7. Plates were allowed to sit for 10 minutes and then inverted and stored in the incubator until the next lab.
8. Lysate was harvested from the two flooded plates and filtered with a 0.22 um syringe filter into a tube.
9. The workstation was cleaned using aseptic technique and materials were properly stored and disposed of.

Observations:

• PA 3 had many bubbles
• 2 plates flooded, 4 plaque assays made

Interpretations/Next Steps:
The procedure was complete. The next step will be to flood the plates and harvest their lysate, and then calculate the titer.

Serial Dilution Plates and Control: