STEWART LABORATORY

 

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In Vitro AdCre INFECTION

 

 

 

Plate the cells at a relatively low density, and the infection should be performed in a minimal amount of media. This protocol is for a 6 well plate,
but you can scale up or down as needed

 

 

1.      Trypsanize cells; quench

 

2.      Spin down the cells and re-suspend in a very small amount of infection media (you can dilute later if necessary); count the cells

 

3.      Dilute with infection media so that the concentration is about 2 x 105 cells/mL

 

4.      For each well, use 500 uL of cells. Add the appropriate amount of virus and mix well (Probably around 1-5 L of high-titer stock you want to get an MOI of around 10)

 

5.      Plate cells and incubate for ~ 2 hrs at 37C

 

6.      Add 2 mL full media and let cells grow for ~ 48 hrs

 

 

 

Infection media: DME + Pen/Strep + 2% IFS + Glutamine

 

Full media: DME + Pen/Strep + 10% IFS + Glutamine