Rationale: In order to guarantee a webbed plate so we can conduct Transmission Electron Microscopy I have received 1mL of Katherine’s lysate and I will calculate the titer and the amount needed to web a plate. I will place more than the calculated required amount so it will definitely be webbed.

Procedure:

1. Counted plaques on plate and took average diameter of plaque.
2. After calculating titer realized that there wasn’t enough lysate to create a webbed plate so plate was flooded.
3. Added 8mL of phage buffer to plate then parafilmed it and stored at 4ºC overnight.

Observations: 514 plaques were formed with 25μL of lysate. This means that the titer is 2.056 * 10^4. The diameter of the plate was 86mm and the average diameter of a plaque was 0.82mm. This means that the amount of pfu needed to web a plate is 1.1*10^4. So the amount of lysate needed to web the plate is 535μL of lysate.

Conclusions and Next Steps: Now I must make a plate with 1070μL of lysate in order to guarantee a webbed plate.