November 7th, 2018
Shepard Saabye

Objectives and Rationale:
Find and amplify the titer of Justin’s lysate in order to mark the phage in TEM.

Previous Results: Cooper and Michael used a 10^-4 titer lysate, and counted 27 plaques on an agar plate that slipped. The plaques were .4 millimeters in radius on average, and the agar that remained on the plate was 35 millimeters in radius. This means the overall titer of the lysate was 2.7*10^7, just shy of a medium titer.

• Procedure:
• Found total number of plaques to web plate (1914),
• Divided plaques needed by the titer, (1914/2.7*10^7) to find the lysate required to make enough plaques to web the plate (7.09 uL of 10^-2)
• Added 8mL LB Broth and 90 uL CaCl to a 50 mL tube
• Collected 3 vials containing .5 mL Arthrobacter
• Added 7.09 uL 10^-2 Lysate to the first vial
• Added 14.18 uL 10^-2 Lysate to the second vial
• Added 2.1 uL 10^-1 Lysate to the third vial
• Added 10 mL 2X TA to the 50 mL tube
• Pipetted 4.5 mL from the 50 mL tube to each vial containing arthro
• Poured all three arthro tubes and the remaining control 50 mL tube to their respective pre-marked plates
• Let sit and placed in incubator at 27 degrees overnight

Analysis and Interpretations: The titer needs to be very high for TEM, and it takes a lot of time to get a high titer. Its a very intensive process to properly web a plate.

Future Plans: Either repeat amplifying or run the TEM again after finding the titer.