Rationale: After a successful plaque assay, the purpose of this experiment is to pick a plaque for purification and perform a plaque assay.

Description of Procedures:

The workstation was cleaned using aseptic technique before beginning the lab. An ethanol burner was lit to create an aseptic work zone as well. To begin, a plaque was chosen to be picked on the plate and labeled p1. The plaque was the picked with a pipette tip and swirled into  phage buffer. It was then vortexed to mix the tube thoroughly. 10 ul of phage buffer solution was then mixed into 0.5 ml of arthrobacter and allowed to sit for 10 minutes. While waiting, the top agar for the plaque assay was made for four plates. 8 ml of LB broth, 90 ul of CaCl2, and 10 ml of 2x Ta were put into a tube and pipetted up and down to mix. Immediately after to ensure that the top agar did not solidify, 4.5 ml of the TA solution was added to the arthrobacter and poured onto a plate labeled LIP 10-1-18 Picked Plaque 1- PA. 4.5 ml were also poured onto a control plate labeled SS EAG LIP 10-1-18 Control. The plates were then allowed to sit for 10 minutes before being stored in the incubator until the next lab. While waiting for the plates to solidify, it was realized that the correct amount of phage buffer was not aliquotted for this experiment. The picking process was then repeated with the same plaque and the correct 100 ul of phage buffer. It was labeled LP P1 and stored in the refrigerator until the next lab when a plaque assay will be performed. When finished, the workstation was cleaned using aseptic technique and the materials were properly stored and disposed of.

Observations/Results:

• There were many plaques found on the last plaque assay
• Bubbles were observed on the control plate.
• The phage buffer was not aliquotted correctly.

Interpretations/Next Steps:

The procedure was completed incorrectly. The wrong amount of phage buffer was used, so the plate made is very diluted and will most likely have no plaques. The plaque was re-picked and stored in the refrigerator until the next lab. The next step will be to run a new plaque assay with the picked plaque.

Plaques found