3/13/13
- Attempted ligation following the same protocol outlined on 3/4/13.
- Grew four liquid cultures of KAH182, following the same procedure (pick a colony and place into LB broth), in case this ligation attempt fails.
> Ligations
|
1 |
2 |
|
Insert DNA (KAH182) |
3.0 |
--- |
|
Vector DNA (KAH201) |
0.5 |
0.5 |
|
2x lgn buf (Roche) |
5.0 |
5.0 |
|
T4 ligase (NEB) |
1.0 |
1.0 |
|
dH2O |
0.5 |
3.5 |
|
|
10 μL |
10 μL |
|
- Follow quick transformation protocol
- Add 30 uL DH5α-Turbo to each ligation; plate on 100 μg/mL Amp agar
> Ligations (corrected version, 3/04/13)
Ligation |
Plate results (lig : neg crtl) 03/13/13
|
1. KAH201 (S/P)/3403, 25 ng + KAH182 (X/P)/1675, 26 ng |
0
|
2. KAH201 (S/P)/3403, 25 ng |
0
|
|