Neon Transfection Conditions

Link to Life Technology's Neon Transfection System

Optimization Conditions

Transfection Conditions

Unless noted, transfections were carried out with the following conditions:

  • Cells were grown in Advanced RPMI + 4% FBS
  • Transfection mix was approx. 1ug pEGFP-C3 DNA (~4.7kb) per 240,000 cells
  • Transfection was carried out in 10ul tips, and cells grown in 24-well plate following electroporation
  • FL was measured at 24hr and 48hr following transfection

Transfection Results

Results were determined by FACS, measuring green fluorescent signal. The viable cells gate was set based on the forward/side scatter dot plot, capturing the viable cell population. Two FL gates were set based on the FL histogram looking at only viable cells: (1) M1, set so that <1.0% of the untransfected cells fell within the M1 gate, and then going all the way to the end of the FL plot (from the far right of the histogram, to the end). (2) M2, set arbitrarly at 10^2 FL units, to capture the more highly fluorescent cells. Numbers summarized below are % total viability, and then calculated % total cells that are viable and fall in the FL gate.



Transfection done with approx. 1.8ug pEGFP-C3 DNA (~4.7kb) per 220,000 cells




Transfection done with approx. 1.8ug pEGFP-C3 DNA (~4.7kb) per 225,000 cells