I'm making reprogramming of MEFs experiment and I subcultured the cells twice then I used Piggy bac, Sleeping beauty and episomal transposition systems to compare between the efficiency of each using electroporation(300v) to get the plasmids in the cells with the helper plasmids, after 4 days of electroporation i used IPs medium and on the 9th day I stained the cells with AP staining ,but I didn't found positive staining cells ,so I repeated the electroporation with lower voltage pulse (250v) and at the same conditions stained it but this time nearly all the cells washed away, I don't know what happened and why no positive staining happened or no cells in the second trial?!!
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