Hello everyone,
I wanted to ask if anyone has been able to successfully transfect HEK293 cells with the BioRad Gene Pulser XCell and the pre-set settings used for it. I've often ended up with cells that do not express our GFP reporter associated with proper transfection.
I've been using the recommended 1*10^6/ml concentration of cells in gene pulser buffer and 10 ug/mL of plasmid DNA.
I also wanted to ask what the efficiency rate of transfection has been post-electroporation, as well as the time it takes you to transfer cells from the cuvette to a plate of fresh, warm media. Thank you!
--Cheers,
Serena
Kousalya Lavudi
You can perform HEK293 transfection by electroporation (125V and 2.5PL). make 2 groups. 1. 1 million cells and EV in 100ul of optimum media. 2. 1 million cells and desired plasmid (1-2ug) concentration is suggestible. check the omega value (desirable is less than or equal to 0.05). After the pulse, immediately add 100ul of media and transfer all 200ul into the dish (3ml media is desirable) as fast as you can. check the GFP expression after 36 hours.
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