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Stable Transfection in Sf9 cells


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#1 eagerscientist

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Posted 10 February 2005 - 01:19 PM

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

#2 expresson_help

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Posted 11 February 2005 - 01:41 AM

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.

#3 eagerscientist

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Posted 11 February 2005 - 05:48 AM

thanks




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