Rationale: Two plates flooded and the flooded lysate made was used to perform serial dilutions to calculate titer.

Procedure: Plates containing 30µL lysate and 10µL lysate, which were flooded 11/13, were filtered under a vacuum hood. Filtered lysate placed in a 15ml vial labeled Flooded lysate x7 original lysate. 10µL lysate transferred to microcentrifuge cap with 90µL PB. The solution was diluted down to 10^-4. Plaques assays were then created to help count titer. The following recipe was used for the experiment (three plates for experiment and one for the control):

• 2mL LB broth (x4)
• 2.5mL 2X TA (x4)
• 22.5µL CaCl2 (x4)
• Lysate
  • 10µL 10^-2 lysate
  • 10µL 10^-3 lysate
  • 10µL 10^-4 lysate

In a test tube, o.5mL Arthrobacter phage was added with lysate from the above experiment (three total lysates), and 4.5mL of the solution made from the above recipe were added to the test tube. Test tube solution poured onto a plate, which sat for 15 minutes, and then placed in the incubator at 27 degrees Celsius for 24 hours.

Observations: Control from 11/12 experiment was contaminated. Switched LB broth solutions and 2X TA solutions for new ones. New solutions were used for this experiment.

Conclusions: Titer will be calculated if the plate yields countable plaques. If not, amplification will continue via revisiting lysate x7. Amplification will continue until high titer is obtained.