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TransMessenger Transfection Reagent Handbook

orter assay
problem Include positive controls to ensure that the reporter assay is working properly. Serum Serum may contain RNases. We therefore recommend transfection in the absence of
serum. Excessive
cell death
Excessive exposure
of cells to
TransMessengerRNA
complexes If excessive cell death is observed after 3 h incubation, reduce the time of cellcomplex
incubation to 12 h. In addition, wash particularly sensitive cells (e.g., primary cells)
with 1.52 volumes of medium without serum instead of PBS after removal of the
TransMessengerRNA complexes. Concentration of
TransMessengerRNA
complexes is too high
Cells are stressed If cell death is still excessive after reducing the exposure time, decrease the amount of TransMessengerRNA complexes added to cells. Avoid stressing cells with temperature shifts and long periods without medium during washing steps. It is particularly important for RNA transfection that the cells are in good condition. Therefore, ensure that cell density is not too low at transfection. For adherent cells, the optimal confluency for transfection is 8090% (for singlestranded RNA) or 5080% (for siRNA). Be sure to seed cells a minimum of 24 h before transfection. Poor RNA quality RNA used for transfection should be of high quality. Impurities present in R
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