siRNA / miRNA gene silencing Human - U251 cofilin-1 (CFL1)

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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Found 1 matching solution for this experiment

pcDNA™3.1 (+) Mammalian Expression Vector + Lipofectamine® 2000 Transfection Reagent

pcDNA™3.1 (+) Mammalian Expression Vector

Thermo Fisher Scientific

Lipofectamine® 2000 Transfection Reagent

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
	siRNA1 (5′-AGCGCAAGAAGGCGGUGCUTT-3′), siRNA2 (5′-GAGGAUCUGGUGUUUAUCUTT-3′) and siRNA3 (5′-GGUGUCAUCAAGGUGUUCATT-3′) siRNA2 had a more potent silencing effect compared with that of siRNA1 and siRNA3. Add diluted siRNA to diluted Lipofectamine Reagent   Incubate for 5 min at RT and add to cells.  Replace medium after 6h and re-incubate cells for 3 days at 37°C and analyse cells at 24,48 and 72 h.

Protocol tips
siRNA1 (5′-AGCGCAAGAAGGCGGUGCUTT-3′), siRNA2 (5′-GAGGAUCUGGUGUUUAUCUTT-3′) and siRNA3 (5′-GGUGUCAUCAAGGUGUUCATT-3′) siRNA2 had a more potent silencing effect compared with that of siRNA1 and siRNA3. Add diluted siRNA to diluted Lipofectamine Reagent   Incubate for 5 min at RT and add to cells.  Replace medium after 6h and re-incubate cells for 3 days at 37°C and analyse cells at 24,48 and 72 h.

Protocol tips

siRNA1 (5′-AGCGCAAGAAGGCGGUGCUTT-3′), siRNA2 (5′-GAGGAUCUGGUGUUUAUCUTT-3′) and siRNA3 (5′-GGUGUCAUCAAGGUGUUCATT-3′)

siRNA2 had a more potent silencing effect compared with that of siRNA1 and siRNA3.

Add diluted siRNA to diluted Lipofectamine Reagent  

Incubate for 5 min at RT and add to cells. 

Replace medium after 6h and re-incubate cells for 3 days at 37°C and analyse cells at 24,48 and 72 h.

Manufacturer protocol Publication protocol 1 Relevant paper

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