I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and also I eager to know the transfection rate by lipofectamine reagents and which kind of lipofectamine is the best for this cell line?
Hello Farzane
There are difficulties encountered while transfecting MDA-MB-231 cell line. I used lipofectamine 2000. It did not give me much success (15%). Toxicity is also observed using this reagent. I suggest you go for lipofectamine 3000 which has higher efficiency and much lower toxicity. Lipofectamine 3000 is also better than Fugene HD.
Lipogene reagent from Genemedi is another reagent which you can try. I have not used lipogene.
Regards.
First, try to use QIAGEN Effectene. Second, try to transfect when the cells are almost confluent (80%), so that their doubling overnight would give you 100% confluence.
Confluence is the key.
Hello !
I have been using these cells for my work and I have to say that they are hard to transfect. Lipofectamine was expensive for me, so I used PEI.
I grow cells to 70-80% confluency. Working plasmid dose was 8ug/100mm dish for me. Plasmid to PEI ratio was 1:3. I diluted plasmid and PEI in 1ml optimem or serum-free DMEM in seperate tubes and incubated for 5 min at RT. Then, I added PEI solution to plasmid solution and incubated for 20 min at RT.
Btw, I always pre-heat PEI to 37C and mix well by vortexing before use. It really helped me increasing transfection efficiency.
Good luck
@ Malcolm Nobre
Hello Malcolm
Thanks a lot for your good answer
did u hear about DNAfectamin?
@ Yulia Panina
Thanks for your advice
If u want to have a competition between Effectene and lipofectamin 3000 for mda mb 231 transfection which one u choose?
how about their efficiency rate?
@ Miray Turk
Hello Dear Miray
Thanx for your comprehensive answer
how much the efficiency rate was with this protocol?
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