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Team:DTU-Denmark/Notebook/27 August 2013
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(→PCR reactions) |
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| Line 25: | Line 25: | ||
| HF buffer || 10 || 10 | | HF buffer || 10 || 10 | ||
|- | |- | ||
| - | | MilliQ || | + | | MilliQ || 31.5 || 27 |
|- | |- | ||
| - | | FW primer || 3 | + | | FW primer || 3 || 3 |
|- | |- | ||
| - | | RV primer || 3 | + | | RV primer || 3 || 3 |
|- | |- | ||
| - | | template || 1 | + | | template || 1 || 1 |
|- | |- | ||
| DMSO || - || 2.5 | | DMSO || - || 2.5 | ||
Revision as of 17:49, 27 August 2013
27 August 2013
Contents |
lab 208
Main purpose
Who was in the lab
Procedure
PCR reactions
| compound | amount (in uL) | |
|---|---|---|
| standard mix | additive mix | |
| dNTPs | 1 | 1 |
| X7 ploymerase | 0.5 | 0.5 |
| HF buffer | 10 | 10 |
| MilliQ | 31.5 | 27 |
| FW primer | 3 | 3 |
| RV primer | 3 | 3 |
| template | 1 | 1 |
| DMSO | - | 2.5 |
| 50 mM MgCl2 | - | 2 |
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 | - |
Results
Gel
- 1 kb ladder
- pSB1C3 for Biobricks, HF buffer
- pSB1C3 for Biobricks, HF buffer, 5%DMSO
- pSB1C3 for Biobricks, GC buffer
- pSB1C3 for Biobricks, GC buffer, 5%DMSO
- negative pSB1C3 for Biobricks
- pZA21::ara for USER ligation with Nir
- pZA21::ara for USER ligation with Nir (duplicate)
- 1 kb ladder
Note: It seems the ladder overflew.
Conclusion
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