Rationale

New titer and web calculations were made since titer is not high enough. Made more webbed plates for new calculations for titer.

Detailed Procedures

1. Took 0.5 mL Arthro and added 10 microliters of 10^0. Sat for 15 mins.
2. Took 4 mL of LB Broth, 5 mL 2X TA, and 45 microliters of 1 M CaCl2 into a 15 mL vial.
3. Took 4.5 mL of top agar mixture and poured onto a plate labeled as “control.”
4. Took rest of 4.5 mL top agar mixture and poured into a 15 mL vial labeled as “10 microliters.”
5. Took mixture from step 1 and added to 15 mL vial labeled as “10 microliters.”
6. Took “10 microliter” vial and poured onto “10 microliters” plate.
7. Let both plates solidify for around 15 mins and inverted into incubator.

Results

Lab Day 31 Current titer= 1.1 x 10^8

Lab Day 31 web= 0.2627 microliter of 10^0

Conclusion

Plates from lab day 30 did not have any contamination. Plaque size compare to webbed plates from a while ago seems to get smaller. Since pipettes can’t measure 0.2627 microliters, used 10 microliters instead. Since it is the last day, flooding and serial dilutions cannot be made and continue DNA extraction, TEM, or nanodrop. This phage was from a donor, results will be looked over by the donor and compare results.

plaque size is small, used a microscope and found 11 pfu on 10^-5 spot

newly webbed plates and control