I am looking for a protocol for plasmid transfection into MCF-10A cells.
At the moment we are using Lipofectamine LTX Plus, but the transfection efficiency is low and most of the cells die.
Do you have any useful tips on how to reduce the toxicity of the transfection or a different transfection reagent that works fine?
Have you tried PEI (polyethyleneimine) ? I would recommend a PEI:DNA ratio of 3:1, but titrate the ratio to obtain optimal transfection efficiency and minimal toxicity.
Have you ever used Electrical transfection instrument(Life Technologies Neon™)?for the common cell lines, it will possess higher transfection efficiency and lower toxicity.
Thank you for the answers. I tested some titrations of Lipofectamine LTX, and a lower LTX concentration seems to do the trick. But if it doesn't work, I will try PEI or electrical transfection!
After 2 years of working with MCF10A cells I came to the conclusion that PEI works the best. The transfection efficiency is still low, but its better than all the other transfection reagents and less stressful than nucleofection. And it has the nice benefit that it is really cheap :)
or with a PEI-derived transfectant avoiding toxic effects of PEI alone...
https://viromer-transfection.com/wp-content/uploads/MCF-10A-transfection-viromer.pdf
Hi,
I am planning to do some transient transfection experiments on MCF-10A.
Víctor González-Tarragó
- So, sir as you recommended above, can you please share your transfection protocol using lipofetamine 3000.- Badges
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