07/25/11
- ✓ Transfection: for real-time microscopy
Transfections: miR-luc sensors, 196/MV8 set
> Lipofectamine LTX
> Follow Jason's protocol, 12-well plates
Plate 2
| Wells |
Cells |
Plasmid / treatment |
3 μg DNA = |
|
| 1 |
U2OS |
183-DF4/MV8 |
5 μL
|
| 2 |
U2OS |
183-DF5/MV8 |
5 μL
|
| 3 |
U2OS |
183-DF6/MV8 |
4 μL
|
| 4 |
U2OS |
183-177/MV8 |
4 μL
|
| 5 |
JDS33-47 |
176/MV8 |
1.3 μL
|
| 6 |
JDS33-47 |
176/MV8 +dox |
1.3 μL
|
| 7 |
JDS33-47 |
177/MV8 |
1.8 μL
|
| 8 |
JDS33-47 |
177/MV8 +dox |
1.8 μL
|
| 9 |
JDS33-47 |
183/MV8 |
5 μL
|
| 10 |
JDS33-47 |
183/MV8 +dox |
5 μL
|
| 11 |
JDS33-47 |
183-177 |
4 μL
|
| 12 |
JDS33-47 |
183-177 +dox |
4 μL
|
> 2 hours prior to microscopy (10 am)
Lipo LTX
Note: set up 3x vol reactions
> Dilute 1 μg DNA in dH2O (10 μL final)
> Add 190 μL Opti-MEM to 10 μL DNA
> Add 1 μL PLUS reagent --> R.T/ 5 min.
> Add 3 μL Lipo LTX --> R.T/ 30 min.
> Add ~200 μL complexes (drop-wise) to each well (1 ml med. each)
> Added transfection complexes at 11 am.
> Real-time microscopy started at 1 pm (48 hrs, 2 hr time points)
| 
Project name
Main project page
Previous entry Next entry
