I am trying to replace the TAP medium with N-free TAP medium after the CW15 has been cultured in TAP medium for 3 days with centrifuge at 1500 g for 5 min followed by pipetting sever time and resuspended into N-free TAP medium. However, the cell been crushed seriously after the replacement operation finished (more than half? I haven’s calculate the crush ratio.). I am confused what's happened with it for the previous experiment went well. Can anyone give me some advice?
I will try centrifuge at lower speed (around 800 g or less for 2 min). When the cells are full of lipid bodies, they are more fragile.
It's my great honour to have your answer, Prof. Li-Beisson.
I will follow your suggestion and feed back soon.
Reply again:
Dear. prof. Li-Beisson, the cell crush ratio decreased since we followed your comment. However, we found another reason (caused by our negligence) should be responsible for the serious crush ratio. Thank you very much.
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