"
September/9 September 2009
From 2009.igem.org
1.colony check
( sample No.) (no. of colonies)
9 50<
24 0
29 0
30 0
31 8
32 2
9 1
2.digestion and ligation
digestion with restriction enzyme
K204024
| Vector | Insert | ||
|---|---|---|---|
| terminater(1-23L) | 6 | No. 14-A | 12 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 4 |
| total | 20uL | total | 20uL |
K204029
| Vector | Insert | ||
|---|---|---|---|
| 2-18F | 6 | 2-16 | 10 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 6 |
| total | 20uL | total | 20uL |
K204030
| Vector | Insert | ||
|---|---|---|---|
| 2-18F | 10 | 1-19B | 6 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 10 |
| total | 20uL | total | 20uL |
↓
37degree 3hr
↓
Agarose Gel Electro-phoresis
No. 24 , 29 , 30 , 10-D , 10-E , ladder
↓
16 degree O/N
3.Min prep
Result of Nanodrop concentration check
(sample No.) (concentration)
1-1D 27.5
10-D 85.1
10-E 117.2
4.transformation
ligation sample No. 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40A ,40B sample No. 24 , 29 , 30
5.digestion and ligation
digestion with restriction enzyme
K204041
| Vector | Insert | ||
|---|---|---|---|
| 1-1D | 12 | 1-12M | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 4 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204042
| Vector | Insert | ||
|---|---|---|---|
| 1-1D | 12 | 1-8I | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 4 | dH2O | 8 |
| total | 20uL | total | 20uL |
↓
37degree 3hr
↓
Agarose Gel Electro-phoresis
No. 41 , 42 , ladder
↓
16 degree O/N
