Hello there. I will try to transient transfect the U-2 OS cell line (an osteosarcoma cell line) with miRNA mimics using Lipofectamine RNAiMAX in a 6-well plate format. Does anyone have any experience with transfection with these cell line? Can anyone suggest how much cells per well and/or how much Lipofectamine should I use? Any tips at all?
Thanks in advance.
Hello, I am working for other reagents, the Viromers, which are not lipid-based as Lipofectamines but rather polymer-based nanoparticle reagents. Just to give my expertise, U-2 OS cells are not particurlarly difficult to transfect and our customers use in general our standard protocol without any further optimization. Which means it should also be ok with the standard protocol of RNAiMAX. We recommend to be around 1x10^6 cells/6-well but be careful, it's always better to adjust the cell density empirically for any new experiments. Avoid being at confluence, 70% is okay.
Please, check attached doc for illustration of what our reagents can achieve in this cell line. Best, Sandra
Sandra Lagauzère Thank you very much. We have Viromer® RED and Viromer® YELLOW in our lab, but I don't know if I can perform miRNA transfection with any os these two reagents, as it is used for transfection of plasmid DNA. Can you tell me if one of the two (RED or YELLOW) can be used for miRNA transfection without any issues? Thanks again.
Alan Jhones Barbosa de Assis Thanks for the additional info. Unfortunately, no, the Viromers RED and YELLOW are not optimal for miRNA, you should rather use Viromer BLUE.
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