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September/14 September 2009
From 2009.igem.org
1. colony check
Checked the cell cultures transformed and plated out yesterday for colony formation and made an approximate count of the number of colonies.
(plate number)-(location on plate) (no. of colonies)
1-16I >50
1-1K >50
1-16C >20
1-4K >20
ligation sample
No. 18 >50
No. 41 >50
No. 42 >50
No. 45 0
No. 46 0
2.digestion for ligation check
digestion with restriction enzyme
for sample No. 27 , 28 , 29 , 30 , 35 , 36 , 39
3.DNA extraction
2-24I
4.ligation check and gel purification
UP : for ligation sample No.17 , 37 , 38
DOWN : ligation check No. 27 , 28 , 29 , 30 , 35 , 36 , 39
ligation check : failure?
5.digestion and ligation
sample No. 20 , 29 , 35 , 27 , 28 , 29 , 30 , 36 , 39
K204020
| Vector | Insert | ||
|---|---|---|---|
| 1-6I | 5 | 1-8I | 5 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 11 | dH2O | 11 |
| total | 20uL | total | 20uL |
K204029
| Vector | Insert | ||
|---|---|---|---|
| 2-18F | 6 | 2-16L | 12 |
| EcoRI | 1 | EcoRI | 1 |
| XbaI | 1 | SpeI | 1 |
| No.2 | 2 | No.2 | 2 |
| dH2O | 10 | dH2O | 4 |
| total | 20uL | total | 20uL |
K204035
| Vector | Insert | ||
|---|---|---|---|
| 1-2M | 5 | 1-14L | 12 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 11 | dH2O | 4 |
| total | 20uL | total | 20uL |
K204027
| Vector | Insert | ||
|---|---|---|---|
| 1-6I | 3 | No. 19 | 9 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 13 | dH2O | 7 |
| total | 20uL | total | 20uL |
K204028
| Vector | Insert | ||
|---|---|---|---|
| 2-18H | 3 | 2-16N | 8 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 13 | dH2O | 8 |
| total | 20uL | total | 20uL |
K204030
| Vector | Insert | ||
|---|---|---|---|
| 2-18F | 2 | 1-19B | 5 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 14 | dH2O | 11 |
| total | 20uL | total | 20uL |
K204036
| Vector | Insert | ||
|---|---|---|---|
| 1-14N | 10 | 1-12M | 2 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 14 |
| total | 20uL | total | 20uL |
K204039
| Vector | Insert | ||
|---|---|---|---|
| 1-14N | 10 | 2-8E | 3 |
| SpeI | 1 | XbaI | 1 |
| PstI | 1 | PstI | 1 |
| No.2 Buffer | 2 | No.2 | 2 |
| dH2O | 6 | dH2O | 13 |
| total | 20uL | total | 20uL |
6.transformation
sample No. 14 , 17 , 41 , 48 , 2-24I , 45 , 46 , 1-14N
