Cells have been
passaged too
many times Cells that have been passaged a large number of times tend to change their growth behavior, morphology, and transfectability. When cells with high passage numbers are used for replicate experiments, decreased transfection efficiencies may be observed in later experiments. We recommend using cells with a low passage number (<50 splitting cycles). No or very small
gene silencing
effect observed
after siRNA
transfection Design of siRNA
sub-optimal The design of an siRNA can have a large effect on its gene silencing efficiency. QIAGEN offers expert advice on siRNA design for maximum gene silencing effect. For more information visit www.qiagen.com/sirna. Incubation time
post-transfection too
short The gene silencing effect observed on the protein level is dependent on a proteins expression level and its rate of turnover within the cell. Perform a time course experiment to determine the optimal time point for analysis. Problems with
experimental design RNAi effects may not be seen for some genes targeted with certain siRNAs in some cell types. If possible, repeat experiments using a different cell type and/or siRNA. Include where possible both positive and negative controls in your experiments. QIAGEN offers siRNA that has been functionally tested for s
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