I would like to know which volumes You use to split a 100 % confluent 150 mm COS-1 cells dish in order to obtain an 80% confluent 150 mm dish 24 h after splting. Which volumes of tripsine do You use and with how many ml of DMEM++ do you block the reaction? And then, with which volumes do You work afterwards to seed in new 150 mm dishes for a transfection pourpouse.
There is another product called Accutase. This does detach the cells but not harmful to the cells. After this you dont have to neutrilize it.
The volume of trypsin does not matter, use just enough to remove cells from the dish, about 1-1.5 ml after first washing with PBS. Growth rates vary a bit depending on media and serum concentrations. Volumes of media do not matter, you need 30-40 for a 150mm dish. What matters is how much of the confluent plate you transfer to a new plate. I would try 1:2 or 1:3..
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