07/27/11
- Transfection: miR-luc sensors "198/MV8" set (9) and CFP-only controls (9)
'Transfections: miR-luc sensors, 198/MV8 set', CFP-only conrols
> Lipofectamine LTX
> Follow Jason's protocol, 12-well plates
Plate 2
| Wells |
Plasmid (MV8 vector) |
3 μg DNA = |
|
| 1-3 |
186-198 |
19 μL
|
| 4-6 |
184-198 |
26 μL
|
| 7-9 |
187-198 |
27 μL
|
| 10-12 |
188-198 |
17 μL
|
| 13-15 |
189-198 |
26 μL
|
| 16-18 |
190-198 |
26 μL
|
| 19-21 |
191-198 |
22 μL
|
| 22-24 |
183-198 |
22 μL
|
| 25-27 |
192-198 |
22 μL
|
| 28-30 |
186 |
12 μL
|
| 31-33 |
184 |
17 μL
|
| 34-36 |
187 |
12 μL
|
| 37-39 |
188 |
14 μL
|
| 40-42 |
189 |
11 μL
|
| 43-45 |
190 |
14 μL
|
| 46-48 |
191 |
12 μL
|
| 49-51 |
183 |
14 μL
|
| 52-54 |
192 |
12 μL
|
| 55-57 |
no DNA |
n/a
|
Lipo LTX
Note: set up 3x vol reactions
> Dilute 1 μg DNA in dH2O (10 μL final); 3x = 3 μg DNA in dH2O (30 μL final)
> Add 190 μL Opti-MEM to 10 μL DNA; 3x = 570 μL Opti-MEM, 30 μL DNA
> Add 1 μL PLUS reagent --> R.T/ 5 min.; 3x = 3 μL PLUS
> Add 3 μL Lipo LTX --> R.T/ 30 min.; 3x = 9 μL Lipo LTX
> Add ~200 μL complexes (drop-wise) to each well (1 ml med. each); Grow cells at 37°C
> Flow cytometry tomorrow (7/28/11)
| 
Project name
