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TransMessenger Transfection Reagent Handbook

lows transfection efficiency to be easily followed.

Designing an siRNA
The design of an siRNA is a critical factor in its ability to mediate gene-specific silencing. QIAGEN provides an siRNA oligonucleotide synthesis service that offers expert advice on the design of an siRNA to achieve optimal gene-silencing effects. For more information visit www.qiagen.com/sirna.

Measuring the gene-silencing effect
The gene-silencing effect can be monitored at the protein level by western blotting, immunofluorescence, gene functional analysis, or FACS. Silencing can also be monitored at the mRNA level by real-time RT-PCR. QIAGEN offers the QuantiTect SYBR Green RT-PCR Kit and the QuantiTect Probe RT-PCR Kit for highly sensitive and specific two-step and one-step real-time RT-PCR analysis. All expression data should be compared to levels of a housekeeping gene to exclude the possibility of non-specific effects.

General recommendations
The guidelines and recommendations below are based on RNAi experiments performed in 24-well plates. Please keep in mind that these are only guidelines, and the efficiency of transfection can be dependent on many different parameters, such as siRNA quality, cell type, passage number, and confluency of the cells at the time of transfection. Variations of these parameters should be considered if varying the amounts and ratios of siRNA and TransMessenger Reagent used does not lead to the desired results. If you would like to discuss optimization of your conditions please contact our Technical Services Department (see inside front cover).

RNAi information online
The QIAGEN Transfection Tools
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