When picking single clones, I use cloning cylinders. A problem which I have faced is that when I use 100 uL of trypsin per cylinder, no cells adhere when directly transferred to a 48-well plate. I have inferred that this is because there is too much trypsin per unit media in the well. What is the optimal way to transfer from a cloning cylinder? Should I move the cells into a larger well? Or just use less trypsin?
Hi,
You could add 100 ul of trypsin , remove immediately, let the cells incubate for few miutes, look under the microscope to insure that the cells have detached , add between 100 to 200 ul of complete media to neutralize the trypsin and transfer to the 48 well plate. I think that should help you.
Thanks
100 ul is enough, just check your cells under microscope to make sure that you have detached them.
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