I have applied Lipofectamine 3000 and Versatile DNA/siRNA transfection reagent jetPRIME® to transfect PX459 plasmid into A549 and NCI-H1975, all the operations are performed according to the manufacturer's instruction. But 24h after transfection, more than half of the cells died, Moreover, after adding 1ug/ml purinomycin, most of the cells died within two days, and finally the monoclone could not be screen. I'm not sure if it is a problem with my cell activity or something else, here is a picture of my H1975 cells before transfection.
In such scenarios, it would be beneficial to pause and conduct some quality checks.
Thank you for sharing the image of your cells pre-transfection. However, could you kindly provide pictures of the mock control as well?
Moreover, I would suggest including an additional fluorescent control to demonstrate the efficacy of transfection.
This data will assist us in comprehensively examining the situation and evaluating any potential cytotoxicity concerns.
Best wishes!
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