siRNA / RNAi /miRNA transfection Rat - H9c2 (myoblast) Cationic and neutral lipids

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.

Start discussion

No discussions found

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

Protocol tips
Always add your plasmid to the buffer before adding Xfect Polymer.

dilute 5 µg of your plasmid DNA with Xfect Reaction Buffer to a final volume of 100 µl.

Incubate for 10 min at room temperature.

Add the nanoparticle complex solution dropwise to the cell culture medium.

Incubate the plate at 37°C for 4 hr to overnight.
TurboFect Transfection Reagents

Thermo Fisher Scientific

Upstream tips
Seed ~5 × 10^4 cells

Protocol tips
Dilute 1µg of DNA in 100µL of serum-free DMEM or other serum-free growth medium.

Incubate 15-20 minutes at room temperature.

Add transfection reagent/DNA mixture to samples and incubate at 37°C in a CO2 incubator.
Downstream tips
Analyse expression after 24-48 hours
Protocol tips
Dilute siRNA in culture medium without serum.

Incubate the samples for 5–10 min at room temperature
Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!
Become shareholder Discussions About us Contact Privacy Terms