siRNA / RNAi /miRNA transfection Rat - A-10 Cationic lipid based

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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Found 3 matching solutions for this experiment

Oligofectamine™ Transfection Reagent

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream tips
Do not add antibiotics to media during transfection; this causes cell death.	Mix all the complex reactions and incubate the cells at 37°C in a CO2 incubator for 4 hours. Add 50 µL of growth medium containing 3X the normal concentration of serum without removing the transfection mixture and assay for gene activity at 24–72 hours post-transfection.

Upstream tips
Do not add antibiotics to media during transfection; this causes cell death.
Protocol tips
Mix all the complex reactions and incubate the cells at 37°C in a CO2 incubator for 4 hours. Add 50 µL of growth medium containing 3X the normal concentration of serum without removing the transfection mixture and assay for gene activity at 24–72 hours post-transfection.

Manufacturer protocol Publication protocol 1 Relevant paper

TurboFect Transfection Reagents

Thermo Fisher Scientific

Upstream tips	Protocol tips	Downstream 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.	Analyse expression after 24-48 hours

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

Manufacturer protocol Publication protocol 1 Relevant paper

HiPerFect Transfection Reagent

Qiagen

Upstream tips	Protocol tips	Downstream tips
	Dilute siRNA in culture medium without serum. Incubate the samples for 5–10 min at room temperature

Protocol tips
Dilute siRNA in culture medium without serum. Incubate the samples for 5–10 min at room temperature

Manufacturer protocol Publication protocol 1 Relevant paper

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