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Team:Heidelberg/Templates/DelH week24
From 2013.igem.org
23-09 - 29-09-13
Generation of DelH plasmids pFS02, pFS03, pFS04 and pFS05
Amplification of pFS_04 Backbone (FS_85 to FS_94; 4.1 kbp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| pSB6A1 | 1 |
| FS_85: (1/10) | 1 |
| FS_94: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
Amplification of pFS_02 Backbone (HM_17 to FS_78; 4.1 kbp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| pSB6A1 | 1 |
| HM_17: (1/10) | 1 |
| FS_78: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
Amplification of pFS_02 Backbone (HM_17 to FS_85; 4.1 kbp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| pSB6A1 | 1 |
| HM_17: (1/10) | 1 |
| FS_85: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
Amplification of pFS_03 Backbone (FS_86 to FS_87; 4.1 kbp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| pSB4K5 | 1 |
| FS_86: (1/10) | 1 |
| FS_87: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
Amplification of DelH fragment (FS_84 to FS_66; 4.1 kbp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| D. acidovorans SPH-1 | 1 |
| FS_86: (1/10) | 1 |
| FS_87: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
Amplification of ccdB fragment for pFS_04 (FS_90 to FS_93; 700 bp)
- Reaction
| Reagent | Amount [µl] |
|---|---|
| pDonor-Plasmid | 1 |
| FS_90: (1/10) | 1 |
| FS_93: (1/10) | 1 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 35 | 98 | 1 |
| 72 | 1:25 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
