It's weird because sometimes it works fantastically, but one day suddenly the transfection process kills every single cell. Any tips for working with PEI/
What PEI do you use- is it the linear polymer or the branched polymer? what Concentration of PEI is your stock solution? What is the final concentration of PEI in your culture? What cells are you transfecting?
You have provided too little information.
Refer to Boussif et al., (http://www.pnas.org/content/92/16/7297.full.pdf)- This is the first description of PEI as a transfection tool.
Polyethylenimine, Linear (MW 25,000) from Polyscience Europe. I'm using 3ug PEI per 1ug of DNA. Right now I'm transfecting 293T cells in 100mm with 10ug DNA.
Thanks in advance!
With different PEI batches, it is possible that the efficacy and toxicity would be different. I would try titrating the amount of PEI per ug of DNA that you use.
Also, the amount of DNA that goes into the transfection has a great effect on cell viability. You will be able to achieve good transfection efficiency with just 1ug of DNA in a 10cm dish. You could try 2 or 3 ug.. If you want to be more thorough, you could titrate the total amount of DNA added to the transfection mix and find your optimal transfection efficiency.
Yes, definitely you are right. That should have been my first shot. I used to transfect 1ug in six well plates, and I thought that 5ug wouldn't be enought to 100mm plate. I have to start from scratch.
Thanks!
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