2 replies to this topic

[Pranita]

I am trying to use Cellular Lights transfection kit for actin-GFP. Can any one suggest me how about optimising the transfection protocol?Actually reagent is quite expensive and therefore I want to use minimum required amout for the opimisation.

[powertoeki]

I will do n=2, duplicates at least... According to Invitrogen Cellular Light transfection kit for ex actin-GFP, the standard protocol is for 100mm plate with 1-4  x 10^56 cells per plate.   You can use  smaller plates to reduce all the trasfection reagent used in a ratio of surface area or vol. capacity.  I usually adjust using vol.capacity. Also, using minimal transfection medium will help to save on reagents. hehe.. :-)

Happy transfection~

[bob1]

Optimise the ratio of DNA:transfection reagent, usually do 6:1, 3:1, 1:3, 1:6, and 2:3

Optimise the amount of DNA transfected. For one well of a 6 well plate 1 ug is usually fine, but up to 5 ug may be used.