Transfection of T47D cells with FuGene in 10cm plates - (Apr/17/2007 )
I am wanting to transfect a 10cm (or even just 6cm) plate of T47D cells using FuGene6, however I have been struggling to get it to work.
I have successfully transfected T47Ds in 24 well plates (using 500ng of DNA and a 3:1 Fugene6:DNA ratio), however I just can't seem to figure out the amount of DNA required to transfect a larger plate.
Does anyone have the conditions worked out for this?
-Nay-
QUOTE (Nay @ Apr 18 2007, 04:51 AM)
I am wanting to transfect a 10cm (or even just 6cm) plate of T47D cells using FuGene6, however I have been struggling to get it to work.
I have successfully transfected T47Ds in 24 well plates (using 500ng of DNA and a 3:1 Fugene6:DNA ratio), however I just can't seem to figure out the amount of DNA required to transfect a larger plate.
Does anyone have the conditions worked out for this?
I have successfully transfected T47Ds in 24 well plates (using 500ng of DNA and a 3:1 Fugene6:DNA ratio), however I just can't seem to figure out the amount of DNA required to transfect a larger plate.
Does anyone have the conditions worked out for this?
more relevant as plate size is volume of medium and cell number and grade of confluency; if your transfection in 24 well was successful, just upscale your conditions to the larger well sizes with corresponding volume of medium and cell number
-The Bearer-
Multiply your amount of cells, DNA, Fugene etc with the difference in size between 6 cm and 24-well plate. I've scaled up/down transfections (with other cells) between 10 cm and 24-well, no big deal.
-vairus-