BHK cells transfection - (Jun/12/2007 )
hi there
does anybody know what should be the concentration of the DNA to transfect BHK cells
i tried using 5 microgram of DNA using lipofectamine but I got very very low transfection efficiency
not even 10%
please help me
thanks
-neuroscientist-
What size of dish/plate are you using? 2-4 ug for a 6 well plate or 35 mm is reasonable range to use. Have you tested different DNA to lipofectamine ratio? I would suggest you testing this parameter on smaller size multi-well plates (48 well-24 well) to conserve reagent and DNA. BHK21 is very easy to transfect and you should get 50 + % positive with little problem. Just watch for possible toxicity issue related to the transfection porcess.
-genehunter-1-
QUOTE (genehunter-1 @ Jun 12 2007, 07:56 AM)
What size of dish/plate are you using? 2-4 ug for a 6 well plate or 35 mm is reasonable range to use. Have you tested different DNA to lipofectamine ratio? I would suggest you testing this parameter on smaller size multi-well plates (48 well-24 well) to conserve reagent and DNA. BHK21 is very easy to transfect and you should get 50 + % positive with little problem. Just watch for possible toxicity issue related to the transfection porcess.
As genehunter suggested, you need to optimise your transfection.
Try out different DNA amounts and different lipofectamine volumes. Also try different cell densities.
-scolix-