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TransMessenger Transfection Reagent Handbook

n. Incubation time is determined by the assay and RNA used.
Note: Compared to DNA transfection, optimal expression levels may be obtained earlier following transfection with RNA. Cells transfected with a chloramphenicol acetyltransferase (CAT) reporter RNA are typically incubated for approximately 24 h post-transfection to obtain maximal levels of protein expression. Table 3. Starting points for optimizing the transfection of adherent cells in different formats * We recommend using medium without serum or antibiotics to avoid possible RNase contamination.
If transfections are performed in 96- or 48-well plates, dilute TransMessenger Transfection Reagent with Buffer EC-R to a total volume of 10 l or 20 l, respectively, before addition to the RNAEnhancer R mixture prepared in step 3.

RNA interference (RNAi), siRNA, and TransMessenger Transfection Reagent

Scientists at QIAGEN have successfully performed RNAi experiments using TransMessenger Transfection Reagent and a gene-specific siRNA (1). Below are some guidelines based on information from current literature (25), and our own experiences.

Controls for RNAi experiments
Controls should always be included in RNAi experiments. An siRNA with a scrambled oligonucleotide sequence can be used as a negative control. Library siRNAs that are directed against human gene products and have been shown to deliver gene-specific silencing in RNAi experiments are available from the QIAGEN siRNA Oligonucleotide Synthesis Service. Fluorescently labeled siRNA al
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