Fugene 6 transfection efficiency? - (Jan/09/2008 )
Can I know the DNA to fugene ratios you are using with your 9 to 12kb plasmids? so that I can start from there.
I have used for our ~10kb plasmids, 3:1 ratio (fugene:DNA) for a single well in a 6 well plate.
1:3 for the most time, but sometimes I use 1:4 as well for different cell types. We've never use it on 293T, coz almost anything on our hands works great on it. Its like to shot a foot soldier with a missile. :-)
Maybe you can increase the cells to 80-90% confluent. Sometimes, incubating DNA-Fugene time to 45 minutes may improve the efficiency. Good luck!