I am planning to use HiPerfect transfection reagent from Qiagen for siRNA transfection in XPC SV40 transformed fibroblasts (adherent cells). Can anyone please share their experience in using this reagent for siRNA transfection.
Hello Lavanya
I work with Hiperfect transfection, but used in functional studies for miRNAs in human monocytes and macrophages. However, miRNAs or siRNA transfection depedent on conditions in your experiments. I advice you try different concentration of your siRNA eg. 50, 25, 12,5 and 2.5nM. In other to know its effect over cell phenotype or viability (Bax/Bcl2 ratio by PCR or anexin - PI).
Best regards
You can optimize your system (its dependent on the cell line and incubation time) and I got good result in 10nM and 50nM siRNA concentrations.
I'm also using the Hiperfect transfection reagent for siRNA transfection in melanoma cells. However, it seems that I don't get transfected cells? I tried siRNA concentrations ranging from 7.5 to 50 nM and it just doesn't work. I assess the efficiency with the Cell Death positive control from Qiagen but the cells just al stay alive.
I was wondering how much of the transfection reagent you add to form complexes with?
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