Hi everyone,
I am currently working on optimizing transfection protocols for HEK293 cells using nucleofection. I'm aiming to achieve the highest possible transfection efficiency. Could anyone recommend a nucleofection program - other than CM-130- that has worked well for them with HEK293 cells? We have a 4D-Nucleofector X Unit - Lonza in the lab. Additionally, any tips or adjustments to the standard protocols that might help improve efficiency would be greatly appreciated.
Thanks in advance!
Not sure why you're considering nucleofection with hek cells. Transfection efficiency of these cells with most transfection reagents approaches 100%...and is likely less strssful.
Hi Sébastien. Thank you for the reply. I’m testing the efficiency of some guides that I designed and base editors. Because I work with Epidermolysis Bullosa, which is a rare disease, and patient cells are very difficult to be obtained, I’m using HEK to standardize the protocol and proceed with the experiments. I appreciate any information you may provide.
Nucleofection (per se) will not be readily extrapolatable to primary cells. That being said, for optimizing the gRNA etc.. I would prioritize HEK and transfection reagents (cheaper, higher throughput). Then you'll have the difficult task of working with primary cells (where nucleofection or transduction will likely be needed).
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