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Team:Heidelberg/Templates/Del week16 AG
From 2013.igem.org
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==12-08-2013== | ==12-08-2013== | ||
===Amplification from FS_02 to FS_11; 22.8 kb=== | ===Amplification from FS_02 to FS_11; 22.8 kb=== | ||
Latest revision as of 13:32, 1 October 2013
Contents |
12-08-2013
Amplification from FS_02 to FS_11; 22.8 kb
_71.8_71.7_71.4_71.0_70.5_70.0_69.5_69.0_68.5_68.1_67.8_67.7besch.png)
- Reaction
| Reagent | DelAG |
|---|---|
| Template | D.acidovorans SPH-1 colony |
| Primer fw | 4.5 µl FS_02 |
| Primer rev | 4.5 µl FS_11 |
| DMSO | 1 µl |
| Phusion Ready Mix | 10 µl |
- Conditions
| Biometra TProfessional Basic | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 72.0 - 67.0 (ΔT = 0.5) ↓ 0.5 | 5 | |
| 72 | 7:20 | |
| 18 | 98 | 1 |
| 70.0 - 65.0 (ΔT = 0.5) | 5 | |
| 72 | 7:20 | |
| 1 | 72 | 10min |
| 1 | 10 | inf |
Results:
- none of the amplifications led to yields of DNA sufficient for Gibson Assembly, therefore the used combination of primers seems not to be sufficient for amplification of 22.0 kbp
13-08-2013
Amplification from FS_02 to FS_11_short; 22.8 kb
- Reaction
| Reagent | DelAG |
|---|---|
| Template | D.acidovorans SPH-1 colony |
| Primer fw | 4.0 µl FS_02 |
| Primer rev | 4.0 µl FS_11_short |
| DMSO | 2 µl |
| Phusion Ready Mix | 10 µl |
- Conditions
| Biometra TProfessional Basic | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 72.0 - 67.0 (ΔT = 0.5) ↓ 0.5 | 5 | |
| 72 | 7:20 | |
| 18 | 98 | 1 |
| 70.0 - 65.0 (ΔT = 0.5) | 5 | |
| 72 | 7:20 | |
| 1 | 72 | 10min |
| 1 | 10 | inf |
