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Team:DTU-Denmark/Notebook/27 August 2013
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{| class="wikitable" style="text-align: right" | {| class="wikitable" style="text-align: right" | ||
| - | ! compound !! amount | + | ! compound !! amount (in uL) |
|- | |- | ||
| || standard mix || additive mix | | || standard mix || additive mix | ||
|- | |- | ||
| - | | | + | | dNTPs || 1 || 1 |
| + | |- | ||
| + | | X7 ploymerase || 0.5 || 0.5 | ||
| + | |- | ||
| + | | HF buffer || 10 || 10 | ||
| + | |- | ||
| + | | MilliQ || 18 uL | ||
|- | |- | ||
| FW primer || 3 uL | | FW primer || 3 uL | ||
| Line 27: | Line 33: | ||
| template || 1 uL | | template || 1 uL | ||
|- | |- | ||
| - | | | + | | DMSO || - || 2.5 |
| + | |- | ||
| + | | 50 mM MgCl2 || - || 2 | ||
|- | |- | ||
|} | |} | ||
Revision as of 17:48, 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 | 18 uL | |
| FW primer | 3 uL | |
| RV primer | 3 uL | |
| template | 1 uL | |
| 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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