The following cell line specific conditions are provided as a starting point for transfecting wild-type cells with TrueGuide Synthetic gRNA and TrueCut Cas9 Protein v2 using the Neon Transfection System 10 μL Kit. Further optimization of the electroporation or nucleofection conditions may be necessary for best results.

Cell typeSourceMediaNumber of cells/10-μL reaction (× 103)TrueCut Cas9 Protein v2/gRNA (ng/pmoles)Neon electroporation conditions*
Well format24-well
HEK293Human embryonic kidneyDMEM1501250/7.5 1150 V/20 ms/2 pulses
U2OSHuman osteosarcomaMcCoy5A1501250/7.5 1400 V/15 ms/4 pulses
A549Human epithelial lung carcinomaDMEM1201250/7.5 1200 V/20 ms/4 pulses
THP1Human peripheral blood monocyte leukemiaRPMI2002000/12 1700 V/20 ms/1 pulse (#5)
K562Human leukemia bone marrowRPMI2001250/7.5 1700 V/20 ms/1 pulse (#5)
iPSCHuman induced pluripotent stem cellsEssential 8801500/10 1200 V/20 ms/2 pulses (#14)
iPSCHuman induced pluripotent stem cellsStemFlex801500/10 1200 V/30 ms/1 pulse (#7)
Human primary T-cellHealthy donor derivedOpTmizer + 2% human serum2001250/7.5 1600 V/10 ms/3 pulses (#24)
Jurkat T-cellHuman peripheral blood lymphocyteRPMI2001250/7.5 1700 V/20 ms/1 pulse (#5)
HepG2Human hepatocellular carcinomaDMEM1201250/7.5 1300 V/30 ms/1 pulse (#8)
N2AMouse brain neuroblastomaDMEM1001250/7.5 1400 V/30 ms/1 pulse (#9)
*Recommendations for the Neon electroporation settings are based on the culture conditions tested.