Rationale: Flooded a plate and added both flooded PB together and ran a plaque assay

Procedure:

• Aseptic zone to prevent contamination
• Obtained webbed plate from incubation and added 4mL of PB to the plate
• Allowed the plate to swirl / sit on a shaker table for a hour and a half
• After the allotted time, obtained a syringe filter and a 22μL filter tip and filtered PB into a 25mL vial
• Obtained a 50mL conical vial and three plates
• Added 10μL lysate into 0.5mL arthrobacter and 20μL lysate into another 0.5mL arthrobacter
• Pipetted 6mL LB Broth and 67.5μL CaCl2 into the 50mL vial
• Added 7.5mL 2XTA into the 50mL conical vial and immediately aded 4.5mL into the arthrobacter + lysate vial(s) (Respectively) and immediately plated
• Allowed the plates to sit for 15 minutes and then moved to incubation

Observations:

• The plate was almost throughly lysed; there was very little lawn of bacteria without plaques
• Added a team members’ flooded PB into the PB from the previous webbed plate to obtain a total of 8mL of flooded filtered lysate

Next Steps/Conclusion: Will be checking the status of the plaque assay next lab. If positive, will be calculating the new strength of the titer and will determine if another webbed plate will be needed to continue raising the strength of the titer