I'm making reprogramming of MEFs experiment and I used Piggy bac, Sleeping beauty and episomal transposition systems.
After electroporation i froze the cells, then i thawed the cells but there was many died cells in some plates and in some was like like pieces of cells floating .
then when i wanted to repeat it with the same conditions the MEFs after electroporation was not attaching to the plate or barely there is some attached.
so what you could advice me to do?
Generally speaking, It is not a good idea to immediately freeze the cells after electroporation. Since the cell membrane was already permeabilized due to electroporation, freezing them would cause additional damage.
Give the cells sufficient time to come to normal condition by incubating them overnight and only if necessary, freeze them the next day, otherwise screen your cells for successful expression of your target.
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