I recently transduced cells in a 6 well plate by spin infection. 1 mL of medium and 100 uL of concentrated lentivirus was put in the well. However, after I spin the plate with a speed of 1500 rpm X 50 min, I found half of the cells dead due to the cells were not covered by medium during the centrifugation. I tried different models of centrifuge including Thermo Legend X1R and Eppendorf 5810R, none of them works. Increasing the medium to 2 mL didn't help, and I saw protocols online used 1 ~ 1.5 mL medium for 6-well plates. Anyone has the same issue or any ideas?
It looks like you are too much spining the cells. Try reducing the time! 5 minutes should be okay. Long duration will definitely kill the cells! After 5 minutes you should incubate your cells.
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