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Team:DTU-Denmark/Notebook/25 August 2013
From 2013.igem.org
25 August 2013
Contents |
Lab 208
Main purpose
- colony PCR
Who was in the lab
Kristian, Henrike
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
Conclusion
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