Rationale

Today we will calculate the titer of our lysate from previously performed spot titer test. After calculating the titer, we will web 5 plates.

Procedure

• Established an aseptic zone.
• The calculated titer was used to determine the amount of lysate needed to web a plate.
• 26.7 µL of 10^-2 lysate was added to 0.5 mL of Arthrobacter. This process was repeated 5 times and each mixture was set aside for 10 minutes.
• 12 mL of LB broth, 135 µL of CaCl2, and 15 mL of TA was added to another vial.
• 4.5 mL of the mixture in the second tube was combined with each vial and was poured onto a plate. Five plates were produced along with a control.
• The plates were set aside for 10 minutes to solidify. After 10 minutes, the plates were placed inside an incubator.

Observations

High titer calculations:

Spot Titer Test:

The 10^-4 lysate dilution was used in calculations due to the 10^-3, 10^-2, 10^-1 lysate dilutions creating webbed or lysed spots.

Conclusions/Next Steps

Next, we will flood our hopefully webbed plates to produce a high titer lysate.