siRNA / miRNA gene silencing Human - PANC-1 DNMT1/3b

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

s4215

Thermo Fisher Scientific

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
Seed 1 × 10^5 cells per well	The DNMT1 siRNA sequences were 5′-GGAUGAGAAGAGACGUAGAtt-3′ and 5′-UCUACGUCUCUUCUCAUCCtg-3′, and the DNMT3B siRNA sequences were 5′-GCUCGUCUCCUAUCGAAAAtt-3′ and 5′-UUUUCGAUAGGAGACGAGCtt-3 Final concentrations - 7 nM 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 2 days at 37°C.

Upstream tips
Seed 1 × 10^5 cells per well
Protocol tips
The DNMT1 siRNA sequences were 5′-GGAUGAGAAGAGACGUAGAtt-3′ and 5′-UCUACGUCUCUUCUCAUCCtg-3′, and the DNMT3B siRNA sequences were 5′-GCUCGUCUCCUAUCGAAAAtt-3′ and 5′-UUUUCGAUAGGAGACGAGCtt-3 Final concentrations - 7 nM 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 2 days at 37°C.

Manufacturer protocol Publication protocol 1 Relevant paper

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