Hello everyone,
I am trying to make a CRISPR-Cas9 knockout on lung cancer primary cells using the RNP complex. I am using the Neon Transfection System but so far it failed - all the cells that took the complex were dead. The settings I tried were: puls voltage 1200 v, width 30 ms, pulse number 2.
I am planning to use their optimization protocol on the 24-well plate but maybe any of you have already tried a similar edition on any type of primary cancer cells and can share the settings used?
Hello Joanna!
Did you finished the optimization? Can you share what are the results?
Sure. The settings working the best for me were 1400 voltage, 20 pulse width and pulse number 2. The other one that was working quite fine was 990/40/1 (better transfection but higher mortality) and 1200/40/1 (worse transfection rate but higher viability). Hope you find it helpful!
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