"
September/7 September 2009
From 2009.igem.org
1.ligation
16degree 3hr
↓
transformation
sample No.27 , 28 , 29 , 30 , 1-1D , 9 , 10-D,E , 24
37degree O/N
2.ligation check
for yesterday sample
23-A,B , vector , 22 , 25-A,B,C,D , vector , ladder ligaton OK
3.Min prep
result of nano drop
sample No. conc.
5 14.9 ng/uL
18 14.6 ng/uL
26 11.1 ng/uL
4.digestion and ligation
digestion with restriction enzyme
K204009
| Vector | Insert | ||
|---|---|---|---|
| 1-23L | 8 | No.4 | 8 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204024
| Vector | Insert | ||
|---|---|---|---|
| 1-23L | 6 | No.14 | 15 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 12 | dH2O | 1 |
| total | 20uL | total | 20uL |
K204031
| Vector | Insert | ||
|---|---|---|---|
| 1-6I | 6 | No.5 | 12 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 4 |
| total | 20uL | total | 20uL |
K204032
| Vector | Insert | ||
|---|---|---|---|
| No.26 | 10 | 1-16G | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 8 |
| total | 20uL | total | 20uL |
↓
37degree , 3hr
Agarose Gel Electro-phoresis
No.9-I,V , No.24-I,V , No.31-I,V , No.32-I,V
gel cut
purification
ligation
