11/7 Titer Calculations
Rationale: Calculate the amount of lysate needed to completely web a plate based on the experiments performed on 11/5.
Procedure: Before the experiment was performed, the workspace was cleaned with both Cidecon and 70% Ethanol. Calculated titer with the equation below:
Determined the titer of the experiment performed, so calculations for how much lysate was needed to web a plate shown below:
In a 50mL vial, 2xTA solution was made for the experiment using the equation below:
- 2mL LB Booth (x4)
- 22.5 microliters of Calcium Chloride (x4)
- 2.5mL 2X TA (x4)
- 500 microliters of Arthro (x3)
- 10^-2 lysate
- 7.09µL plate 1
- 14.18µL plate 2
- 2.127 of lysate 10^-1 plate 3
4.5mL of 2XTA solution was added into a test tube containing 10^-2 lysate + Arthrobacter phage solution and quickly poured onto a plate. Plates sat for 15 minutes to solidify, and then inverted and placed in the incubator at 27 degrees Celcius.
Observations: Plates from 11/5 experiments did not sit all the way causing plaques to tear and the TA solution did not completely cover the plate. The control from the experiment performed on 11/5 was positive.
Conclusions: Calculate pfu/µL on 11/12 to determine high/medium titer. If the experiment performed 11/7 is a high titer, flood the plate, and if the titer is medium, redo experiment, same as 11/7 experiment.