Rationale: To make a plaque assay with the positive lysate to receive a high titer

Procedure:

• The tables were thoroughly wiped down with CiDecon and ethanol and an aseptic zone was also set up to prevent contamination.
• 125 µL of positive lysate and 0.5 mL Arthrobacter were placed in a new tube and left alone for 10 minutes to allow for infection.
• Next, 22.5 µL CaCl2 was combined with 2.0 mL of LB broth.
• After 10 minutes, lysate and Arthrobacter were added with LB broth and CaCl2.
• Then, 2.5 mL 2X Top Agar was added to the LB broth and CaCl2.
• The mixture was poured over the plate and then left alone to solidify before being placed in the incubator.

Results and Analysis:

Conclusion:

Due to the contamination of the negative control, a plaque assay was done. First, lysate and Arthrobacter were combined and left alone for 10 minutes to infect. Then, LB broth was added along with the calcium chloride. After the allotted time, the lysate and Arthrobacter were added to the LB broth and calcium chloride. 2X Top Agar was then added to the solution and poured onto the agar plate. For 15 minutes, the plates remained near an aseptic zone to solidify. After, the plates were placed in an incubator.

Future Plans:

When a plate is high titered with no contamination, the plate will be flooded with phage buffer and filtered to perform dilutions.