Rationale:  The purpose of this lab is to collect lysate and perform a spot titer test to determine if the lysate is a high titer.

Description of Procedures:

1. The workstation was cleaned using aseptic technique and an ethanol burner was it.
2. Lysate was harvested from the four flooded plates using a syringe and was filtered into a tube with a 0.22 um syringe filter. Approximately 25 ml were obtained.
3. Serial dilutions were performed out to 10^-5 with the lysate collected(lysate 2) and lysate 1.
4. Top agar was made for one plate, using 2 ml of LB broth, 22.5 ul of CaCl2, 2.5 ml of 2x TA, and 0.5 ml of arthrobacter. The top agar was poured onto a plate labeled LIP 11-16-18 Spot Titer Test. The plate was divided into 7 sections, one for the phage bufer test, and 3 for each of the lysates, 10^-3 to 10^-5.
5. 10 ul of lysate was spotted into each of the six sections, and 10 ul of phage buffer was spotted into its section.
6. The plate was allowed to sit for 15 minutes and then inverted and stored in the incubator until the next lab.
7. The workstation was cleaned and the materials were properly stored or disposed of.

Observations:

• Few bubbles seen on the plate
• Top agar control was made on a second plate by a different group.

Interpretations/Next Steps:
The procedure was complete. The next step will be to determine the titer of the lysate collected and create a TEM grid.

Picture of Plates: