MOI Calculation

Protocol

1. Seed 293T cells at 20,000 cells per well on a 96-well plate 1 day prior to infection.

2. Create a dilution of the virus stock in DMEM with 10% FBS

Example dilution for 1 column of 1 plate:

Row	DMEM	Added
1	30 uL	15 uL Virus stock
2	30 uL	10 uL Row 1 solution
…	…	…
8	30 uL	10 uL Row 7 solution

3. Add 30 uL of diluted virus + 70 uL DMEM with 10% FBS to each well

4. Record how many uL of the original virus stock was used in each well (can be calculated with this excel file)

5. Count and record the cells in 1 well of the plate at the time of infection

6. 24 hours later, analyze the cells via flow cytometry

Note

Rather than seeding the cells 1 day before infection, cells can instead be infected in suspension to precisely control the number of cells mixed with the virus. In this case, it is convenient to combine diluted virus with media to a volume of 50 uL in each well, then use the multichannel to add 50 uL cells to each well of the plate.