Rationale

Created 10^-5 dilution to make a newly webbed plate in hope of flooding and creating a high titer.

Detailed Procedure

1. 10 µL of “SJ 11/12/18 FS lysate 10⁰” was added with 90 µL of phage buffer to create 10^-1 dilution which was vortexed.
2. 10 µL of 10^-1 was added with 90 µL of phage buffer to create 10^-2 dilution which was vortexed.
3. 10 µL of 10^-2 was added with 90 µL of phage buffer to create 10^-3 dilution which was vortexed.
4. 10 µL of 10^-3 was added with 90 µL of phage buffer to create 10^-4 dilution which was vortexed.
5. 10 µL of 10^-4 was added with 90 µL of phage buffer to create 10^-5 dilution which was vortexed.
6. Took 98 µL of 10^-5 into 0.5 mL arthro. Set for 15 mins to infect.
7. Took 4 mL of LB Broth, 5 mL of 2X TA, and 45 µL of 1M CaCl2 into 15 mL vial.
8. Took 4.5 mL of top agar mixture and poured onto control plate.
9. Took rest of top agar mixture and combined with arthro and 10^-5 mixture.
10. Took resulting solution and poured onto 10^-5 plate.
11. Allowed both plates to sit for 15 mins.

Conclusion/Next Steps

Since the calculations were 97.6 µL of 10^-5 , only 98 µL were pipetted for easier pipetting. By next lab day, hopefully a webbed plate will be able to flood and make new plates to calculate the new titer. By Friday, hopefully a high titer will be calculated.