Rationale: After flooding the plate, the lysate will be filtered then used to perform serial dilutions to conduct a spot test

Procedure:

• The lysate from the flooded plate was taken using a syringe then filtered.
• Into eight tubes, 90 µL of phage buffer was added to each.
• From the 10^0 dilution, 10 µL of lysate was added to the tube containing phage buffer.
• From the 10^-1 dilution, 10 µL was taken and added to the 10^-2 tube and so on.
• After completing the dilutions, a spot test was made by combining 2.0 mL of LB Broth, 22.5 µL of calcium chloride, and 0.5 mL of Arthrobacter
• Then, 2.5 mL of 1X Top Agar was added then poured onto the plate.
• After 15 minutes of allowing the top agar to solidify, 0.7 µL of each dilution was added to their respective areas within the plate.
• The plate was left alone to allow for the spot dilutions to set in the agar.

Results and Analysis:

While removing the lysate from the flooded plate, the top agar tore.

Conclusion:

After setting up the correct measures to prevent contamination, eight small tubes were filled with 90 µL each. From the 10^0 lysate, 10 µL was transferred to another tube, creating the 10^-1 dilution. From the 10^-1 dilution,  10 µL was transferred to the 10^-2 tube to create the dilution and so on. Next, the spot test was made by combining LB Broth, 1X Top Agar, calcium chloride, and Arthobacter then poured onto a labeled plate.  After 15 minutes, the lysate was dropped in drops of 7 µL in their designated areas. The plates were left alone for 15 minutes to solidify then placed inverted in the incubator.

Future Plans:

Identify the dilution with the highest titer but not lysed then use that lysate to make a plaque assay to determine the titer of the plate, If it has a high titer, flood the plate.