Rationale: After a negative plaque assay from the flooded lysate, the purpose of this lab is to retest the flooded lysate and pick a plaque for purification. This lab will also retest enriched soil C lysate for further study.

Description of Procedures:

1. The workstation was cleaned using aseptic technique and an aseptic zone was made with an ethanol burner.
2. 100 ul of phage buffer was added to a tube. A plaque was picked and mixed in the buffer. The tube was then vortexed to mix.
3. 10 ul of phage buffer with plaque was added to 0.5 ml of arthrobacter, 100 ul of the flooded lysate was added to 0.5 ml of arthrobacter, and 10 ul of soil C lysate was added to 0.5 ml of arthrobacter, and all were allowed to sit for 10 minutes.
4. 8 ml of LB Broth and 90 ul of CaCl2 were added to a tube to create the top agar solution. 10 ml of 2x TA were added the tube.
5. 4.5 ml of the top agar solution was added to 0.5 ml of arthrobacter for each plate, and the poured directly onto plates labeled LIP 10-24-18 PA SC, LIP 10-24-18 PA-P1, and LIP 10-24-18 PA(FL). The rest of the solution was poured directly onto a plate labeled LIP 10-24-18 Control.
6. The plates were allowed to settle for 10 minutes and then inverted and stored in the incubator until the next lab.
7. The workstation was cleaned using aseptic technique and materials were properly stored and disposed of.

Observations:

• 100 ul of flooded lysate was added instead of the usual 10 ul.
• Soil C lysate was retested.
• Some bubbles were seen on the plates.

Interpretations/Next Steps:
The procedure was complete. The next step will be to check for plaques on all the plates, and to flood the plate from the picked plaque.