siRNA / RNAi /miRNA transfection Rat - IEC-6 (Intestinal Epithelial cells) Lipofectamine

The RNA interference (RNAi) is used to inhibit gene expression or translation, by neutralizing targeted mRNA molecules. Two types of RNA molecules such as microRNA (miRNA) and small interfering RNA (siRNA) play a central role in RNAi. Few points have to considered to increase the transfection efficiency of siRNA. Always use healthy, actively dividing cells to maximize transfection efficiency. The confluency of cells should be between 50-70%. Always use the most appropriate siRNA concentration to avoid off-target effects and unwanted toxic side effects. Positive and negative controls should be used for each and every experiment to determine transfection efficiency.

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Protocol tips
Add diluted siRNA to diluted Lipofectamine Reagent

Incubate for 20 min at RT and add to cells.

Replace medium after 5h with 5% heat-inactivated FBS and re-incubate cells at 37°C for 48 h
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