07/18/11
- ✓ Transfection: miR-luc sensor "177/MV8" set
Transfections: miR-luc sensors, 177/MV8 set
> Lipofectamine LTX
> Follow Jason's protocol, 12-well plates
Plate 2
| Wells |
Plasmid (MV8 vector) |
3 μg DNA = |
|
| 1-3 |
186-177 |
22 μL
|
| 4-6 |
184-177 |
22 μL
|
| 7-9 |
187-177 |
16 μL
|
| 10-12 |
188-177 |
18 μL
|
| 13-15 |
189-177 |
17 μL
|
| 16-18 |
190-177 |
15 μL
|
| 19-21 |
191-177 |
13 μL
|
| 22-24 |
183-177 |
11 μL
|
| 25-27 |
192-177 |
12 μL
|
| 28-30 |
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 (7/19/11)
| 
Project name
Main project page
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