What is the necessary time for lipofectamin to make complex with pre-miRNA oligo - (Mar/19/2013 )
I just transfected some cells with pre-miRNA oligos alone without any marker
The point forgot I did not wait 20minutes after adding lipo to plasmid solution.Just3 to 5 minutes after mixing lipofectamin with pre-mirs
is such matter really affects experiment this time?do cells absorb naked dsRNA in case of not having completely formed lipofectamin particles?
I was really disappointed while doing this assay because I do not see the desired effect on protein after several experiments and I do not have any method in my mind to check the transfection effeciency and thats such a disaster to wait after doing western to see what has happened
The time is important, 3-5 min is probably not long enough. Cells shouldn't take up the RNA without the transfection reagent.