siRNA / miRNA gene silencing Human - LLC PKN3

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

SIHK1738

Sigma-Aldrich

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
Seed 3×105 cells per well	siRNA concentratioin-50 nM PKN3 siRNA were as follows: Sense, 5′-AgA cUu GaG gAc UuC cUg GaC aA-3′ and antisense, 5′-uUg UcC aGg AaG uCc UcA aGu Cu-3′ Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 1–3 days at 37°C.

Upstream tips
Seed 3×105 cells per well
Protocol tips
siRNA concentratioin-50 nM PKN3 siRNA were as follows: Sense, 5′-AgA cUu GaG gAc UuC cUg GaC aA-3′ and antisense, 5′-uUg UcC aGg AaG uCc UcA aGu Cu-3′ Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio).  Incubate for 5 minutes at room temperature and add to cells.  Incubate cells for 1–3 days at 37°C.

Manufacturer protocol Publication protocol 1 Relevant paper

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