November 2

10/26 ~ Filtering a flooded plate and performing a plaque assay

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Rationale: Filtered the flooded plate that has sat for 48 hours tot obtain a new lysate; conducted a plaque assay to calculate/test the titer strength.

 

Procedure:

  • Created an aseptic zone to prevent/reduce bacterial contamination
  • Obtained the previous flooded plate from the refrigerator
  • Obtained a syringe filter (22μL) and filtered the PB into a 15mL conical vial
  • Obtained a 50mL conical vial and added in 4mL LB Broth and 45μL CaCl2
  • Added 20μL lysate to 0.5mL arthrobacter to infect
  • Pipetted 5mL 2xTA into the conical vial and immediately added 4.5mL into the arthrobacter + lysate vial and plated
  • Allowed plates to sit for 15 minutes and then incubated

 

Observations:

The new lysate obtained from filtering the PB of the flooded plate

The results of a plaque assay to observe the strength of the previous titer

10^-1 result of the previous titer

 

Next Steps/Conclusion: After performing this plaque assay, will be calculating the new titer strength and aiming to strengthen the titer strength. If the plate is negative / contaminated, will perform another plaque assay to check plaques and calculate the strength.


Posted November 2, 2018 by justin_yu1 in category Justin Yu

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