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Team:DTU-Denmark/Notebook/13 August 2013
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| - | + | ===SLP screening PCR=== | |
| + | The PCR run with condition E (5% DMSO, 2mM MgCl2 with GC buffer) had the highest flexibility; it yields the expected product at different annealing temperatures: 58.4 C, 59.6, C 60.6 C, 61.9 C, 63.2 C, 64.6 C, 65 C. | ||
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Revision as of 15:16, 13 August 2013
13 August 2013
Contents |
lab 208
Main purpose
Who was in the lab
Kristian, Henrike, Gosia
Procedure
PCR (promoter library)
PCR to amplify USER fragments of the constitutive SPL, the constitutive reference promoter and the arabinose reference promoter.
Used 5% DMSO, 2 uL of mM MgCl2 and GC-buffer.
constitutive SPL
- template: pZA21::RFP
- primers: 52a, 52b1
constitutive reference
- template: pZA21::RFP
- primers: 52a, 52b2
arabinose reference
- template:
- primers: 51a,51b1
program
| temperature | time | cycles | |
|---|---|---|---|
| 98C | 2:00 | - | |
| 98C | 2:00 | - | |
| 98C | 2:00 | - | |
| 98C | 2:00 | - | |
| 98C | 2:00 | - |
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Conclusion
SLP screening PCR
The PCR run with condition E (5% DMSO, 2mM MgCl2 with GC buffer) had the highest flexibility; it yields the expected product at different annealing temperatures: 58.4 C, 59.6, C 60.6 C, 61.9 C, 63.2 C, 64.6 C, 65 C.
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