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Team:Heidelberg/Templates/Del week16 G
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(Created page with "==15-08-2013== ===Amplification from SR_01 to FS_23; 6.2 kb=== [[File:Heidelberg_20130814 2log 5xDelGstandard MasterMixrest.png|150px|thumb| Amplification of DelG; run at 100 V, ...") |
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==15-08-2013== | ==15-08-2013== | ||
===Amplification from SR_01 to FS_23; 6.2 kb=== | ===Amplification from SR_01 to FS_23; 6.2 kb=== | ||
Latest revision as of 13:46, 1 October 2013
Contents |
15-08-2013
Amplification from SR_01 to FS_23; 6.2 kb
- Reaction (5x20 µl)
| what | µl |
|---|---|
| D. acidovorans SPH-1 colony | 1 |
| SR_01: (1/10) | 2 |
| FS_23: (1/10) | 2 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 3:20 | |
| 18 | 98 | 1 |
| 64 | 5 | |
| 72 | 3:20 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
- Amplification of DelG was successful
- Bands were cut out and DNA purified using QIAquick Gel Extraction Kit
- Amplification will be repeated to increase the concentration for the Gibson Assembly
16-08-2013
Concentration measurement
the fragment was gel purified
| Fragment | Primer | Date PCR | Concentration |
|---|---|---|---|
| DelG | SR01-FS23 | 15-08-2013 | 52.6 ng/µl |
| DelG after nucleotide removal | SR01-FS23 | 15-08-2013 | 30 ng/µl |
Amplification from SR_01 to FS_23; 6.2 kb
- Reaction (5x20 µl)
| what | µl |
|---|---|
| D. acidovorans SPH-1 colony | 1 |
| SR_01: (1/10) | 2 |
| FS_23: (1/10) | 2 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad T100 | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 66 ↓ 0.5 | 5 | |
| 72 | 3:20 | |
| 18 | 98 | 1 |
| 64 | 5 | |
| 72 | 3:20 | |
| 1 | 72 | 10min |
| 1 | 12 | inf |
Results:
- Amplification of DelG was successful
- Bands were cut out and DNA purified using QIAquick Gel Extraction Kit
