mm/dd/yy
- Outline of experiments/ tasks
Task 1
> Line item 1
Reagent |
Volume |
Conc.
|
reagent 1 |
vol 1 |
conc 1
|
reagent 2 |
vol 2 |
conc 2
|
dH2O |
vol 3 |
|
|
total vol |
--> Reaction conditions
|
Lipo transfection
> Details
Wells |
Plasmid |
DNA |
Volume |
Lipo |
Opti-MEM (total)
|
1 |
name |
2 μg |
### μL |
4 μL |
500 μL
|
2 |
name |
" |
### μL |
" |
"
|
3 |
name |
" |
### μL |
" |
"
|
4 |
name |
" |
### μL |
" |
"
|
5 |
name |
" |
### μL |
" |
"
|
6 |
name |
" |
### 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.
> Remove 500 μL medium from each well; Add 500 μL complexes to each well (4 ml med. each); Grow cells at 37°C
> Refresh medium after 5 hours (ab-free)
Fugene transfection
> Details
Wells |
Plasmid |
DNA |
Volume |
Fugene |
Opti-MEM
|
1-6 |
name |
200 ng |
### μL |
1.8 μL |
18.2 μL
|
7-12 |
name |
" |
### μL |
" |
"
|
13-18 |
name |
" |
### μL |
" |
"
|
19-24 |
name |
" |
### μL |
" |
"
|
> Master mixes (4, x6): 10.8 μL Fugene + 109.2 μL Opti-MEM --> R.T/ 5 min.
> Add 6x vol DNA --> R.T./ 20 min.
> Add 20 μL complexes to each well (1 ml med. each); Grow cells at 37°C overnight
|