Rationale

New titer and webbed plate calculation were recorded. The amount calculated (22 microliters of lysate) will be used to make a new webbed plate.

Detailed Procedure

1. Took 22 microliters of 10^0 lysate and added to 0.5 mL arthro. Sat for 15 mins.
2. Took 4 mL LB Broth, 5 mL 2X TA, and 45 microliters of 1M CaCl2 into a 15 mL vial.
3. Took 4.5 of Top Agar mixture into a plate labeled as “SJ control 11/14/18.”
4. Took remaining 4.5 mL of Top Agar mixture and added arthro+lysate mixture.
5. Poured mixture into a plate labeled as “SJ PA 22 microliters 11/14/18.”
6. Allowed plates to solidify for 15 mins and inverted into incubator.

Results

current titer= 2.4 x 10^6

need to web plate= 21.3 microliters, used 22 microliters.

Conclusion/next steps

Plates were sliding, no bubbles formed on the plate. The current titer calculation is nearly the same titer as last time. In the next lab day, hopefully, the plate will be flooded and perform serial dilutions up to 10^-8 and recalculate for a new, high titer

titer calculations were used from 10^-4 spot