Shepard Saabye
November 12th, 2018
SEA PHAGE Lab Journal

Results from the previous lab: One of the plates was close to webbed. The control came out a little contaminated.

Rationale and Objectives: Today the goal was to generate a new lysate, and have it prepared to calculate titer by the next lab. This is to increase the titer high enough to complete TEM.

• Procedure:
• Established Aseptic Zone
• Flooded webbed plate with 5 mL Phage Buffer
• Let sit on moving platform for 1 hour
• Diluted lysate out to 10^-3 of the original lysate
• Obtained 3 vials of .5 mL Arthrobacter
• Added 10 mL LB Broth, 12.5 mL 2X TA and 112.5 uL CaCl to a single 50 mL tube
• Added 50 uL of 10^0 Lysate to the first Arthro tube
• Added 10 uL 10^-2 Lysate to the second tube
• Added 10 uL 10^-3 Lysate to the third tube
• Added 4.5 mL of Agar mixture to each Arthro tube, and plated
• Plated remaining 4.5 mL Agar mixture on the control plate
• Waited 15 min for Top Agar to solidify
• Left plates in the incubator at 24.5 degrees Celsius

Future PLans: use the results from this lab to determine the titer of our lysate, and decide if that is high enough to continue with TEM.