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Team:Heidelberg/Templates/Del week11 AG
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Revision as of 23:31, 29 September 2013
12-07-2013
Amplification from FS_02 to FS_07; 16.4 kb
File:20130712 DelAG.png
PCR for amplification of DelAG (12.07;FS02+FS07); lane6=Ladder log2, lane7=Ladder 1kb base ruler, lane8=DelAG(12.07;FS02+FS_07) run at 100 V, 0.8 % gel (TAE)
- Reaction
| what | µl |
|---|---|
| D. acidovorans | 1 |
| FS_02: (1/10) | 2 |
| FS_07: (1/10) | 2 |
| Phusion flash Master Mix | 10 |
| DMSO | 1 |
| dd H2O | 4 |
- Conditions
| Biorad MyCycler | ||
|---|---|---|
| Cycles | temperature [°C] | Time [s] |
| 1 | 98 | 10 |
| 12 | 98 | 1 |
| 68 ↓ 0.5 | 5 | |
| 72 | 5:30 | |
| 18 | 98 | 1 |
| 66 | 5 | |
| 72 | 5:30 | |
| 1 | 72 | 15min |
| 1 | 12 | inf |
Results:
- only a smear occured, no specific product was amplified
- PCR will be repeated with a lower, constant annealing temperature
