I am trying to transfect SNU449 cell line by GFP plasmids using genepulserX Bio rad electroporator. I wonder if I can use .4 cm cuvette and voltage needed ! and what is the cell size of SNU449 !
I guess you can use 0.4cm cuvette for your purpose. The voltage and the time constant has an inverse relationship for the electroporation. Thus, you can use 10-40msec with 900-400V/cm field strength. The size of normal hepatocyte is around 20-30µ. I guess cell size should not differ much in case of carcinoma, but i am not sure.
If you work up to 1 kV/cm, you can use 0.1-0.4 cm cuvette, then voltage must be 100-400 V respectively. Thus, cuvette must be anyone within field strength 1 kV/cm (e.g. 1000x0.2=200 V, 1000x0.3=300 V etc). In any case cells 20-30µ will fit between the electrodes up to 0.01 cm (only 10V).
Thank you so much. Kindly If I am going to transfect the cells with crispr cas9. The electroporation will make me less amount of the crispr than chemical transfection using lipofectamind as I have limited amount of crispr cas 9 ?
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