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Team:DTU-Denmark/Notebook/13 July 2013
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=208 lab= | =208 lab= | ||
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Latest revision as of 20:40, 16 September 2013
13 July 2013
Contents |
208 lab
Main purpose today
Make gel purifications to enable USER-reactions and to make PCR of the fragments missing.
Who was in the lab
Kristian
Procedure
Purification gel 1,2 and 3
- 1kb ladder
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- HAO
- 1kb ladder
- cycAX
- cycAX
- cycAX
- cycAX
- AMO
- AMO
- AMO
- AMO
- HAO
- HAO
- HAO
- 100 bp ladder, NEB
- RFP
- RFP
- RFP
PCR of AMO
Did PCR on AMO on two programs: 1-3 on 54°C 4-6 on 52°C Both with the annealing time 3 min.
USER-reactions
There was made USER-reactions with all fragments after purification. AMO was purified even though it was only one very faint band; the others were bright band. The following plates where made:
- RFP USER - RFP inside pZA21
- AMO USER - AMO inside pZA21
- HAO USER - HAO inside pZA21
- cycAX USER - cycAX inside pZA21
- Nir USER - Nir inside pZA21
- Neg. USER - pZA21 linearized only
Conclusion
PCR of AMO with USER-primers was not a success we will see if the new PCR have done better. We will evaluate the transformants on Monday with colony-PCR.
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