2 replies to this topic

[eagerscientist]

Hi i need help

i transfected a plasmid i made with the dna of interest.  Pcr analysis shows that everything is okay with the plasmid.

i transfected sf9 cells with this plasmid trying to make stable cell line.  after 72 hours post transfection, i checked for fluorescence using FACS and  fluorescence microscopy.  no detection at all.

Can anyone help.

thanks

[expresson_help]

If you want a stable cell line, try linearising your plasmid prior to transfection. This increases the rate of integration into the host genome.

If you think transfection efficiency is a problem, you could try NeoPhectin (http://www.expresson...neophectin.html) which works well in hard-to-transfect cell types.

[eagerscientist]

thanks