I'm planning to transfect HepG2 and Huh7 with poly I:C (high molecular weight poly IC from Invivogen using lipofectamine 2000 as transfectant reagent. I would like to do a standard curve using different concentrations of poly IC (1, 2.5, 5 and 10 ug/ml) but I don't really know which would be the ratio poly IC : lipofectamine. (I would first mix the two compounds in Optimum and leave them at RT for 25 min as described by Invivogen protocol).
Can I use the same amount of lipofectamine for each poly IC concentration or should I use a ratio of the two, in this case which would be this ratio?
Hello,
Poly I:C works well without any transfection reagent. However, you can check this paper: https://www.spandidos-publications.com/10.3892/mmr.2016.4848?text=fulltext
Poly (I:C) transfection
For poly (I:C) transfection, 1×105 HeLa cells were seeded in a 12-well plate and maintained at 37°C in a 5% CO2 incubator for 15 h. A dose of 2 µl Lipofectamine™ 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) was added to 100 µl of serum-free medium, following which the mixture was incubated at room temperature for 5 min. At the same time, 10 µl poly (I:C), purchased from Sigma-Aldrich (St. Louis, MO, USA) was added to the 100 µl of serum-free medium. Subsequently, the Lipofectamine™ 2000 and poly (I:C) were mixed gently and incubated at room temperature for 20 min. Following incubation, the cell culture medium was replaced with serum-free medium, which was added to each well containing the Lipofectamine™ 2000 and poly (I:C) mixture, and incubated at 37°C for 4 h. Finally, the medium in each well was replaced with a fresh serum-containing medium.
I hope this helps
Best Regards
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