I am doing Dual luciferase assay in a hard to transfect cell line and got these values-
1) Non-transfected- Firefly 93.16(RLU) and Renilla 123.85(RLU), so F/R is 0.75
2) Empty vector transfected- Firefly 1351(RLU) and Renilla 740.97(RLU), so F/R is 1.82
This same experiment , previously I did in a easily transfected cell line, and noticed that the difference between these two values are very high. In case of the difficult transfected cell line, are the values acceptable? If not, please advice me what should I do?
I think the values appear acceptable. In case you'd like to strengthen the difference, I recommend you to co-transfect an additional vector containing a neomycin (or something else) resistance gene that facilitates the selection of the positively transfected cells.
The difference in the value is ok. You can proceed with the experiment
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