06/03/10
- ✓ Transfection: histone methylation reporter
Lipo transfection
> KAH96 [5xGal4 : HSVtkTATA : K : YFP : YFP : NLSstop2 : polyA : HS4 : HS4]
> KAH142 [DPRE : 5xGal4 : HSVtkTATA : K : YFP : YFP : NLSstop2 : polyA : HS4 : HS4]
Wells |
Plasmid |
DNA |
Volume |
Lipo |
Opti-MEM (total)
|
1 (KAH126-1) |
KAH96/ FlpE |
1.5, 0.5 μg |
2.5, 1.0 μL |
4 μL |
500 μL
|
2 (KAH126-1) |
KAH142/ FlpE |
1.5, 0.5 μg |
2.5, 1.0 μL |
" |
"
|
3 (KAH126-1) |
none |
--- |
--- μL |
" |
"
|
4 (FTRx) |
KAH96/ FlpE |
1.5, 0.5 μg |
2.5, 1.0 μL |
4 μL |
500 μL
|
5 (FTRx) |
KAH142/ FlpE |
1.5, 0.5 μg |
2.5, 1.0 μL |
" |
"
|
6 (FTRx) |
none |
--- |
--- μL |
" |
"
|
> Add Lipo to 250 μL Opti-MEM --> R.T/ 5 min.
> Add DNA to 250 μL Opti-MEM
> Add DNA mix to Lipo mix --> R.T./ 20 min.
> Add 500 μL complexes to each well (4 ml med. total each); Grow cells at 37°C
> Refresh medium after 5 hours (ab-free)
|