For an experiment I am required to use a non-enzymatic dissociation buffer on my cells. I lifted 1205Lu melanoma cells off a T75 flask with 2mL of 5mM EDTA in DPBS. However, after centrifugation in a 15mL conical tube at 1000rpm for 3 minutes, my cells did not form a pellet as they would if I were using TryplE. The pellet was extremely small, and cells seemed to be stuck to the walls of the tube and some were still in suspension. I tried centrifuging at 1200rpm for 5 minutes and could not create a better pellet. Does anyone have any recommendations of how to avoid this?
Gloria Marino
Laurence Stuart Dawkins-Hall
Thank you so much! I also read about the BSA wash and I think it's worth a try in conjunction with a significantly longer spin.
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