Rationale:

Passage the bacteriophage another time to further purify the bacteriophage, and to produce a countable plate

Procedure:

• Created an aseptic zone to prevent contamination of bacteria in the plates/experiment
• Picked a plaque from the 9/26 plaque assay (10^0) and pipetted into 100μL PB
• Created a 10^-1 and 10^-2 dilution by adding 90μL PB into two separate micro-centrifuge tubes and adding 10μL of the previous dilution (10^-1 created by adding 10μL 10^0 into 90μL PB, and same with 10^-2)
• Added 10μL of each lysate into 0.5mL of arthobacter
  • Allowed to sit for 15 minutes to infect the arthrobacter
• Obtained a 50mL vial and added 20mL LB Broth and 225μL CaCl2 into the vial (Multiplied recipe by 10 for 10 plates)
• Added 25mL of 2X TA into the vial, and immediately pipetted 4.5mL TA into the arthrobacter+lysate vial, and plated
• Allowed plates to sit for 15 minutes and then incubated

Observations:

• The 10^0 plate had numerous plaques, but many were split
• 10^-1 and 10^-2 had little to no plaques
• The control plate of the group was contaminated

Next Steps/Conclusion:

After this passage, the next step would be to calculate the titer of the lysate. This would be done by counting the plaques and obtaining the measurements of the plate/plaques. The bacteriophage worked with should be considerably purified, since this is the sixth(Or so) purification of the bacteriophage