"
September/23 September 2009
From 2009.igem.org
1. colony check
sample no no. of colony
22 100<
25 100<
40 100<
47 100<
X1-A 50<
X2-B 2
X1-B 3
X3-B 2
X2-A 50<
1-12M 10<
X3-A 10<
37 0
39 0
2.min prep
result of nano drop
sample no conc.
1-2M 42 ng/uL
1-12M 61
X1-B 47
X2-A 98
38 234
X3-B 88
36A 247
36B 23
X2-B 96
X1-A 61
X3-A 64
3.digation
X4-A
| Vector | Insert | ||
|---|---|---|---|
| 32 | 16 | 1-12M | 4 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 0 | dH2O | 12 |
| total | 20uL | total | 20uL |
K204052
| Vector | Insert | ||
|---|---|---|---|
| 1-6I | 10 | 33 | 4 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 10 |
| total | 20uL | total | 20uL |
K204029
| Vector | Insert | ||
|---|---|---|---|
| 2-18F | 8 | 2-16L | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204037
| Vector | Insert | ||
|---|---|---|---|
| 3-1M | 10 | 1-8K | 3 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 13 |
| total | 20uL | total | 20uL |
K204039
| Vector | Insert | ||
|---|---|---|---|
| 3-1M | 8 | 2-8E | 3 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 13 |
| total | 20uL | total | 20uL |
K204046
| Vector | Insert | ||
|---|---|---|---|
| 32 | 13 | 1-8K | 2 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 3 | dH2O | 14 |
| total | 20uL | total | 20uL |
↓
37°C, 2hr
4.trandfromation
sample no 28 , 30 , X2-A , X1-B
5.prepared Lb/Amp plate
6.digation
X5
| Vector | Insert | ||
|---|---|---|---|
| 1-6O | 8 | 1-12M | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
X6
| Vector | Insert | ||
|---|---|---|---|
| 1-12A | 12 | 1-12M | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
X7
| Vector | Insert | ||
|---|---|---|---|
| 1-12C | 8 | 1-12M | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
X8
| Vector | Insert | ||
|---|---|---|---|
| 1-16G | 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 |
↓
37°C, 2hr
gel electrophoresis
gel cut
purification
↓
ligation
7.digation
K204053
| Vector | Insert | ||
|---|---|---|---|
| 3-1M | 8 | 33 | 6 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 | 2 | No.2 | 10 |
| dH2O | 8 | dH2O | 8 |
| total | 20uL | total | 20uL |
7.transfromation
sample no. X4-A , X4-B , 52 29 , 37 , 39 , 46 3-1M , 11
