Co-transfection - (Jul/02/2005 )
If u transfect a mammalian cell line like 293 t cells with 2 seperate plasmids encoding for EGFP and E 47 respectively, what are the chances that the cells will take up only one plasmid instead of both.
-ybbobck-
QUOTE (ybbobck @ Jul 2 2005, 07:32 PM)
If u transfect a mammalian cell line like 293 t cells with 2 seperate plasmids encoding for EGFP and E 47 respectively, what are the chances that the cells will take up only one plasmid instead of both.
I use a method with calcium phosphate to co-transfect 3 plasmids and works so good.
-donisaid-
In general you add so many plasmids (literally 100s of millions to billions) that the limiting factor
is the actual taking up of the plasmid by a cell. Therefore, any cell that takes up one plasmid
will usually (almost always) take up two. Stable transfection is a different matter...
But, if you ever do CAT or luciferase asssay, and normalize with B-GAL, there will almost never be an incidence where a cell takes up one plasmid but not the other.
-mikew-
May I use co-transfection to get the stable cell line expressing 2 genes I am interested?
QUOTE (mikew @ Jul 5 2005, 12:30 PM)
In general you add so many plasmids (literally 100s of millions to billions) that the limiting factor
is the actual taking up of the plasmid by a cell. Therefore, any cell that takes up one plasmid
will usually (almost always) take up two. Stable transfection is a different matter...
But, if you ever do CAT or luciferase asssay, and normalize with B-GAL, there will almost never be an incidence where a cell takes up one plasmid but not the other.
is the actual taking up of the plasmid by a cell. Therefore, any cell that takes up one plasmid
will usually (almost always) take up two. Stable transfection is a different matter...
But, if you ever do CAT or luciferase asssay, and normalize with B-GAL, there will almost never be an incidence where a cell takes up one plasmid but not the other.
-macrosky-
QUOTE
May I use co-transfection to get the stable cell line expressing 2 genes I am interested?
Sure, I've done it. It worked... sort of.
-pBluescript-