Neon electrporation of CRISPR RNPs into MV4-11 cells

Thank you all for participating in this helpful forum. I am wondering if anyone has optimised Neon electroporation (10uL) for human MV4;11 cells for delivery of plasmid DNA and/or Cas9 RNPs and is willing to share their optimal conditions (# cells, plasmid DNA/RNP amount, Volts, pulse time, number of pulses etc.)?


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Purple Bogreabout 4 years ago

I am new and I was surfing a little in this forum. It might be late for you but It could be helpful.
I applied Neon Protocol on MEF fibroblast cells (mouse). They adhere on the plastic of the well. I have no clue about your cell line.

I transfected 1.3uM RNPs (IDT) + 1uM linear dsDNA (4.8kb).

1600V, pulse width: 10ms and pulse No: 3. The number of cells was 54000.

48h of incubation for HDR.

Let me know how it goes..

Good luck

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