Transfection - Optimising Transfection Protocol (Aug/03/2011 )
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.
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.. :-)
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.