hi, i am currently preform the siRNA transfection of ovcars cell with silentfect lipid reagent. kit protocol only said the gene silencing can be monitored at the mRNA or protein levels from 4 to 72hrs after transfection. could someone tell me about what the optimal time of siRNA transfection of ovcar3 cells? thank you~
Jiangnan He In general cells will display low expression of your target protein in 24 hrs span as well. However, in my experience during most of the transfection experiments, 48 hrs is a suitable time.
You can also do a time-dependent experiment and then optimise for yourself what time span suits your experiment. Again transfection of siRNA for 48 hrs should be enough to see transient knock down of your protein.
Hope it helps
As Vikrant Mehta said, I would prefer 48 h transfection time. Start with 50-60 % confluency of cells. Follow the silentfect manufacturer protocol. Dose-dependent and time-dependent experiments can fetch you the exact knockdown efficiency. If you need more details in this regard let me know. Cheers.
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