08/03/10
- ✓ Transfection: Test H3me Reporter clones
- ✓ KAH126-3: expand thawed cells
- ✓ Senescence assay: 6-well plates, -/+ dox, for KAH126-1, 126-3, 132-8, 154-2, FTRx (rotenone, dox, DMSO, u.t.)
- ✓ Cultures: split confluent plates
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
|
| 18 total |
no plasmid |
--- |
0 μL |
1.8 μL |
18.2 μL
|
| 18 total |
KAH59/pcVN |
200 ng |
0.4 μL |
" |
"
|
> Master mixes (20x mix, 2 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
08/04/10
Results: No YFP
08/05/10
Results: Screened for YFP again today and saw lots of YFP+ cells in KAH146-1; re-plate KAH96 & 147 for another transfection
|
Project name
Main project page
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