siRNA / miRNA gene silencing Human - DU145 PLAU

Gene silencing through the use of small interfering RNA (siRNA) has become a primary tool for identifying disease-causing genes. There are several aspects for preparing and delivering effective siRNA to knockdown a target gene. The length of siRNA should be 21–23nt long with G/C content 30–50%. If a validated siRNA sequence for your target gene is not available, use siRNA generated against the entire target gene ORF. Always work with two or three different siRNA constructs to get reliable results. If you are not sure how much siRNA to use for a given experiment, start with a transfection concentration of 10-50 nM and use siRNA-specific transfection reagent to ensure efficient siRNA delivery in a wide range of cells.

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Dharmacon

Bio-Rad Laboratories

Bio-Rad Laboratories

Upstream tips	Protocol tips	Downstream tips
	siRNA concentration-90nM Replace medium 1h before transfection. Add diluted siRNA to siLentFect Reagent to make a mixture and incubate the mixture for 20 min at room temperature. Replace culture medium after 4h of transfection and incubate the plate at 37°C for 24 to 72 h.

Protocol tips
siRNA concentration-90nM Replace medium 1h before transfection. Add diluted siRNA to siLentFect Reagent to make a mixture and incubate the mixture for 20 min at room temperature. Replace culture medium after 4h of transfection and incubate the plate at 37°C for 24 to 72 h.

Manufacturer protocol Publication protocol 1 Relevant paper

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