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Team:DTU-Denmark/Notebook/25 August 2013
From 2013.igem.org
25 August 2013
Contents |
Lab 208
Main purpose
- PCR
Who was in the lab
Procedure
Colony PCR
Performed colony PCR to confirm insert for HAO, AMO, cycAX and Nir transformants. Used Q5 premix with the following reaction mix:
| compound | amount |
|---|---|
| Q5 mix | 25 uL |
| FW primer | 3 uL |
| RV primer | 3 uL |
| template | 1 uL |
| MilliQ | 18 uL |
Template was made by resuspending 1 culture in 100uL MilliQ.
program:
| temperature | time | cycles |
|---|---|---|
| 98C | 10:00 | - |
| 98C | 0:10 | 36 |
| annealing temperature | 0:30 | 36 |
| 72C | 0:20 | 36 |
| 72C | 5:00 | - |
| 10C | hold | - |
details (primers, temp, expected fragment length):
- cycAX - FW_1, RV_2, 61C, 749bp
- HAO - FW_2, RV_3, 64C, 751bp
- AMO - FW_2, RV_3, 63C, 750bp
- Nir - FW_2, RV_3, 64C, 733bp
- Nir - FW_5, RV_6, 71C, 737bp
Results
Conclusion
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