Rationale: Help Justin to obtain a high titter by performing a plaque assay, and alternating some of the variables to obtain a webbed plate.

Procedure: Before the experiment was performed, the workspace was cleaned with both Cidecon and 70% Ethanol. The formula below was used to make the solution for 2 plates (one for the control and one for the plaque assay).

• 2mL LB Booth (x2)
• 22.5 microliters of Calcium Chloride (x2)
• 2.5mL 2X TA (x2)
• 500 microliters of Arthro
• 588µl of Justin’s lysate

2X TA solution was added into a 15mL vial, and in a test tube with 0.5µl Arthrobacter, 588µl of lysate was added. The test tube solution sat for 15 minutes, and then 4.5mL of the 2X TA solution was added to the test tube. The test tube solution was then quickly poured onto a plate, and the rest of the 2X TA solution was poured onto a second plate as a control.

Observations: Gel Electrophoresis performed on 10/29 came back negative. Obtained lysate from Justin to help obtain a high titer. Justin added 488µl of lysate, but in the experiment performed today, 588µl of lysate was added.

10/31 Control

10/31 Plaque Assay

Results: On Monday 11/5/18, calculate how much lysate is needed to obtain how much lysate should be added to obtain a high titer. Check Justin’s flooded plate to help to ensure his plates will have a high titer.