Calcium Phosphate Transient Transfection of NIH/3T3???

Alexander N. Kukushkin kan at mmcd.cyt.ras.spb.ru
Fri Feb 7 11:36:46 EST 1997


>   Hello.
>      Has anyone ever transiently transfected NIH/3T3 fibroblasts by the
>   Calcium Phosphate method?  I have several protocols already, but all of
>   them are for other cell types.  I'm going to be doing Luciferase assays
>   after.
>   Thanks in advance,
>      Roland Halil
>   email:   s635141 at uottawa.ca
>   Neurosciences Research Institute
>   University of Ottawa
>   Ottawa, ON
>
Hi
For long time we perform transient transfection of NIH3T3 cells by the
Calcium Phosphate method using the Gorman's protocol with some modifications
(Gorman C.M., Moffat L.F., Howard B.H. - Mol. Cell. Biol., 1982, 2, 1044).
However, the cell extracts are used for CAT assay. The transfection efficiency
is good according to the data of beta-galactosidase activity as well as result
of the reporter assay.
Alex




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