I am trying to produce stable A549 cell lines targeting my gene knockdown by shRAN. When cells get transfected in next passage they start to die very fast while the cells which are not cloned they remain alive
I think you mean "shRNA", and that may mean the gene you are knocking down is lethal when knocked down.
Try using a control experiment transfecting cells with water (control), and cells with the shRNA. If the control fails to allow the cells to be stable, then the transfection may be causing the lethality.
I completely agree with Carter J Barger. You need to the treatment in same conditions.
Thanks Carter,
I gave a basic approach to these problems, and you added some details, and other possibilities.
However, without knowing your exact protocol, we are shooting darts in the dark. I highly recommend using the logic presented here in your experiments.
Thanks you all for nice comments I found that my gene is lethal
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