08/10/10
- ✓ Transfection: H3me Reporter lines (screen for YFP induction)
- ✓ Microscopy: Fix cells (KAH130, 131 clones)
- ✓ Western: protein preps for KAH130, 131 clones
Fugene transfection
> 12-well plate
Plate 1. KAH96 clones (6 total)
Plate 2. KAH146 clones (6 total)
Plate 3. KAH147 clones (6 total)
Wells |
Plasmid |
DNA |
Volume |
Fugene |
Opti-MEM
|
20 total |
no plasmid |
--- |
0 μL |
1.8 μL |
18.2 μL
|
20 total |
KAH59/pcVN |
200 ng |
0.6 μL |
" |
"
|
20 total |
KAH60/pcVN |
200 ng |
0.4 μL |
" |
"
|
> Master mixes (20x mix, 3 mixes): 36.0 μL Fugene + 364.0 μL Opti-MEM --> R.T/ 5 min.
> Add 20x vol DNA --> R.T./ 20 min.
> Add 20 μL complexes to each well (1 ml med. each); Grow cells at 37°C overnight
|