Rationale:

Perform a plaque assay plate for a adopted lysate to create a high titer plate and flood it to get more lysate. This will be used to create a webbed plate.  

Procedure:  

1. Created a aseptic zone using ethanol burner and cleaned with Cidecon and 70% ethanol.  
2. Obtained 30 uL of adopted lysate and added to 0.5 mL of Arthrobacter.  
3. Waited 10 minutes for to infect.  
4. Obtained a 50 mL tube and and added 2 mL of LB for the plate.  
5. Added 22.5 uL of CaCl2 to the tube as well. 
6. Added 2.5 mL of Top Agar after the 10 minutes were up.  
7. Poured the contents of the 50 mL tube into the Arthro and lysate.  
8. Poured all of it onto a plate.  
9. Let the plate set for 15 minutes until solidified and gels set.  
10. Inverted and placed into incubator to be viewed next class time.  

Observations/ Results: 

The plates created by other members of the group from the past week, created a medium titer plate but not a high titer plate. Also, the plate with more lysate (15 uL) had less phages than the plate with less lysate (14.1 uL).  

There was not enough lysate as well, so flooded the 14.1 uL plate to create more lysate. 

Next Steps/ Conclusion: 

Next class we will observe if the amount of lysate used this class period will create a webbed plate. If there is a webbed plate then we will continue by viewing the plate. If the plate is not webbed then we will try to create a high titer or webbed plate.