I have successfully nucleofected but would like to try transfection. Seems like Lipofectamine 2000 isn't the best bet. Has anyone used another Lipofectamine kit successfully?
I am no expert in this but do have some experience trying different transfecting reagents. My opinion is that you really need to try all possibilities. I am happy with lipofectamine LTX but sometime lipofectamine 200 works better in specific primary cell culture which I can not explain. In my experiment the Lipofectamine 3000 did not work but lipofectamine LTX did. You, and maybe a lot of our peers, may say I am crazy but it is my personal experience. For unknown reason the reverse transfection protocol, which I adpoted from lipofectamine RNAiMax, works better for my experiments but again I do not know why. Now my colleagues in the group use the same method. Any method work for your system is the best method and you have to give it a try. Wish you luck.
I was using the lipophilic reagent Effectene (Qiagen) and it works for me much better then the lipofectamine 2000
Dear Brenna,
we are currently evaluating different transfection reagents available on the market but have not finished the study yet. Unfortunately, most of the reagents cause accumulation of the siRNA in agglomerates in vesicle-like structures within the cell.
When you evaluate a new reagent I recommend using a fluorescently labeled siRNA so that you can analyze the cellular distribution and localization of the siRNA.
Good luck and all the best
Stefan
Why is everyone responding with reference to siRNA/miRNA transfection? How about transfecting THP-1 cells with plasmid DNA? David, what efficiencies do you get with Effectene? How do you keep the THP-1 cells from differentiating after Lipofectamine or electroporation? Apologies for asking so many questions, my experience with modifying monocytes is quite limited. Thank you in advance everyone!
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