Previous Results:

• On 11/8 the plates made of “Lysate #6 10^-2” were flooded. On 11/9 the lysate was collected (Lysate #7) and plaque assay was made by a lab partner. Today, the plate was examined, and although the control did not have any contamination, the assay looked contaminated, as if there were no plaques.

Objective:

• Plaque assays of serial dilutions for Lysate #7 will be redone, due to the results of the last assays. Hopefully a titer can be calculated using these plates

Procedure:

1. Aseptic Zone was created with CiDecon, 70% Ethanol, and an Ethanol burner
2. Lysate #7 was obtained and diluted out to 10^-3
3. Using 90 microliters phage buffer and 10 microliters of lysate in a microcentrifuge tube, the 10^-1 lysate was made. 10 microliters of 10^-1 was transferred into 90 microliters phage buffer to create 10^-2. This was repeated to make 10^-3
4. 10 microliters from each dilution (10^0 – 10^-3) were added to individual tubes of 0.5 mL Arthrobacter. Left to infect for 10 min
5. Overlay Agar was mixed in a 50 mL tube using 10 mL LB broth, 12.5 mL 2x TA, and 112.5 microliters CaCl2
6. 4.5 mL of Overlay was added to each Arthro tube, leaving 4.5 remaining in the 50 mL tube used for control plate
7. All 5 agars were plated, left to harden for 10 min, and incubated for 24 hours

Next Steps:

• During the next lab, the plates will be examined and used to calculate the titer of Lysate #7. Depending on the titer, TEM will be run on the sample