Rationale: Prepared a TEM grid for TEM imaging of the phage

Procedure:

• Obtained the lysate from refrigeration and went down to the TEM lab
• Obtained Uranyless from Aadil and obtained a parafilm strip
• Placed the parafilm strip in a petri dish and pipetted one drop of lysate, two separate drops of water and one drop of uranyless onto the strip (Each ~20μL)
• Using surgical/precision tweezers, picked up a copper grid and placed it shiny side down onto the lysate drop for 5 minutes
  • Following the 5 minutes in the lysate, transferred the grid into the first drop of water and let sit for 2 1/2 minutes; after the allotted time, transferred the grid to the other drop of water and water for 2 1/2 minutes
• After the water bath, transferred the grid into the uranyless and let sit for 1 minute (Time sensitive) and immediately wicked off excess uranyless solution
• Grid was then loaded into the TEM machine and imaged

Observations:

An image of what we believe to be a phage; the size proportions/ratio between the tail and head are off

The parafilm with the 2 drops of water, 1 drop of lysate and 1 drop of uranyless

The grids where the copper grids reside and were transferred out of and into

The completely lysed plate with bacterial growth that came back

Next Steps/Conclusion: There were no discoveries of actual phage; the phage that was found had uncommon ratios between the tail and head. The next steps would be to continue to raise the titer of the lysate to image the phages more easily