06/28/11
- ✓ Transfection: miR sensor test set #1 (no Gal4 binding sites), U2OS plain
Transfections: miR sensor set 1, 0 Gal4 BS
Plate 1: Lipo LTX
> Lipofectamine LTX
> Follow Jason's protocol, 12-well plates
| Wells |
Plasmid (MV8 vector) |
DNA |
Volume |
PLUS reagent |
Lipo |
Opti-MEM (total)
|
| 1-3 |
KAH186 |
1 μg |
5 μL |
1 uL |
3 μL |
200 μL
|
| 4-6 |
KAH184 |
" |
5 μL |
" |
" |
"
|
| 7-9 |
KAH187 |
" |
5 μL |
" |
" |
"
|
| 10-12 |
KAH186-196 |
" |
5 μL |
" |
" |
"
|
| 13-15 |
KAH184-196 |
" |
5 μL |
" |
" |
"
|
| 16-18 |
KAH187-196 |
" |
5 μL |
" |
" |
"
|
| 19-21 |
KAH186-177 |
" |
5 μL |
" |
" |
"
|
| 22-24 |
KAH184-177 |
" |
5 μL |
" |
" |
"
|
| 25-27 |
KAH187-177 |
" |
5 μL |
" |
" |
"
|
| 28-30 |
KAH186-198 |
" |
5 μL |
" |
" |
"
|
| 31-33 |
KAH184-198 |
" |
5 μL |
" |
" |
"
|
| 34-36 |
KAH187-198 |
" |
5 μL |
" |
" |
"
|
--> 37-39. no transfection
Lipo LTX
Note: set up 3x vol reactions
> Add 180 μL Opti-MEM to 5 μL DNA; 3x = 540 μL Opti-MEM, 15 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
| 
Project name
Main project page
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