siRNA / miRNA gene silencing Human - DuCaP SKIL

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

siRNA SKIL

Thermo Fisher Scientific

INTERFERin®

Polyplus transfections

Upstream tips	Protocol tips	Downstream tips
Seed 30000 cells/well	The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) SKIL siRNA concentration- 50 nM Add INTERFERin to siRNA duplexes and incubate for 10min at RT after homogenization. Remove culture medium from cells and add incubate mixture. Incubate the plate at 37°C for 24 to 72 h.

Upstream tips
Seed 30000 cells/well
Protocol tips
The siRNA sequences (5′-to-3′) were: 1: GGCAAGUAAGUCCAUAUCATT (sense) and UGAUAUGGACUUGCCTC (antisense) 2: GGCUCACAGUAGUGGUAATT (sense) and UUACCACUACUGUGAGCCTT (antisense) SKIL siRNA concentration- 50 nM Add INTERFERin to siRNA duplexes and incubate for 10min at RT after homogenization. Remove culture medium from cells and add incubate mixture. Incubate the plate at 37°C for 24 to 72 h.

Manufacturer protocol Publication protocol 1 Relevant paper

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