Rationale: The purpose of this lab is to flood a plate to obtain more lysate to create a webbed plate.

Description of Procedures:

1. The workstation was cleaned using aseptic technique and an ethanol burner was lit to create an aseptic zone.
2. 5 ml of phage buffer was poured onto the plate with plaques.
3. The plate was shaken on an incubator for one hour.
4. While shaking, 4 ml of LB Broth and 45 ul of CaCl2 were added to a tube, to create top agar for two plates.
5. The lysate was harvested from the plate with a syringe and filtered with a 0.22 um syringe filter into a tube. 3 ml of lysate were obtained.
6. 10 ul of the lysate was added to 0.5 ml of arthrobacter and allowed to sit for 10 minutes.
7. 5 ml of 2 x TA were added to the top agar solution and pippetted up and down. 4.5 ml of the solution was added to the 0.5 ml of arthrobacter and poured onto a plate labeled LIP 11-5-18 10 ul titer test. The rest of the top agar solution was poured onto a plate labeled LIP 11-5-18 TA Control.
8. The plates were allowed to sit for 10 minutes and then inverted and stored in the incubator until the next lab.
9. The workstation was cleaned using aspetic technique and materials were properly stored or disposed of.

Observations:

• Plate from last lab was not completely webbed.
• Bubbles on the plates.

Interpretations/Next Steps:
The procedure was complete. The next step will be to calculate the titer and make a webbed plate.

Control, 14 ul Lysate Plate, 15 ul Lysate Plate