November 5th, 2018

Shepard Saabye

Objectives and Rationale: Identify Phage using TEM in a lysate of unknown titer capable of lysing a plate. Identifying this phage will help in the classification of the phage and its characteristics.

Procedure:

• Calibrated TEM
• Allocated Petri Dish and Parafilm
• Placed a strip of parafilm securely inside petri dish top
• Placed a drop of lysate, 2 separate drops of water, and a drop of Uranyl less on the parafilm
• Using precision tweezers, placed a copper grid, shiny side down, on the lysate for 5 minutes
• Moved copper grid to the first drop of water for 2.5 minutes
• Moved copper grid to the second drop of water for 2.5 minutes
• Placed copper grid on uranyl less (UL) for 1 minute, and quickly blotted an excess UL once removed
• Used tweezers to move copper grid into TEM Grid Holder
• Moved Copper Grid to the preliminary Vacuum chamber in the Machine
• Closed Vacuum and observed the grid

Results:

No phages were found.

Analysis and Interpretations:

The titer may have been too low to effectively find phages. In addition, the Uranyl Less may have been ineffective compared to Uranyl Acetate.

Future Plans: We will either repeat the experiment with Uranyl Acetate, or work to increase the titer of the lysate used.