How to Infect Mammalian Cell Cultures with Viruses

  1. Split the cells the day before so the following days the cells are in the desired confluency (40-50% for immunofluorescence studies). Split cell into a glass cover slip if a fluorescence microscope will be made later.
  2. Wash the cells with PBS the following day.
  3. Replace with a new medium containing 0.2% fetal bovine serum.
  4. Add the desired amount of virus at infection (MOI). An MOI of 1 should be sufficient for most studies, but the MOI of about 0.01 to 100 also have been useful to various studies.
  5. Allow the virus binds to the cell surface for 30 minutes to 1 hour (keep the binding constant of time each time) at room temperature in order to synchronize the entry of the virus. (4 degrees incubation on ice may be needed if the virus can enter the cells at room temperature.)
  6. After 30 minutes to 1 hour binding, remove the virus bound to the medium, and replace with new media containing 2% fetal bovine serum.
  7. Place the cell into the 37 degrees C incubator for the desired amount of time. For the new virus protein expressing cells, incubation overnight will usually allow enough time for new proteins to be expressed.

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