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Team:Evry/Notebook/w9
From 2013.igem.org
(Difference between revisions)
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| - | + | 50.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.62 | |
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| - | + | 0.68 | |
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| - | + | 21.1 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.69 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.44 | |
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| - | + | 21.6 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.20 | |
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<td align="center"> | <td align="center"> | ||
| - | + | 0.13 | |
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| - | + | 70.8 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.66 | |
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<td align="center"> | <td align="center"> | ||
| - | + | 0.72 | |
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| - | + | 39.1 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.23 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.40 | |
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| - | + | 25.1 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.37 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.29 | |
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| - | + | 61.1 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.57 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.71 | |
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| - | + | 54.7 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.62 | |
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<td align="center"> | <td align="center"> | ||
| - | + | 0.68 | |
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| - | + | 38.4 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.84 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.78 | |
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| - | + | 114.6 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.66 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.78 | |
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| - | + | 39.2 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.87 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.21 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 77.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.76 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.99 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 78.4 ng/µL | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.61 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.81 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 72.7 | |
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<td align="center"> | <td align="center"> | ||
| - | + | 1.85 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 4.23 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 70.4 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.76 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.16 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 91.5 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.55 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.21 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 65.4 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.70 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.52 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 85.6 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.72 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.65 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 53.5 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.76 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.93 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 77.6 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.69 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.22 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 72.3 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.68 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.22 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 73.9 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.58 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 0.84 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 51.3 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.75 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.10 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 45.9 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.85 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.43 | |
</td> | </td> | ||
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| - | + | 46.1 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.88 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.61 | |
</td> | </td> | ||
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| - | + | 108.3 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.78 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.37 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 67.0 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.88 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.61 | |
</td> | </td> | ||
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| - | + | 103.5 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.87 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.79 | |
</td> | </td> | ||
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| - | + | 28.2 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.06 | |
</td> | </td> | ||
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| - | + | 1.76 | |
</td> | </td> | ||
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| - | + | 29.5 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.95 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.22 | |
</td> | </td> | ||
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| - | + | 70.0 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.90 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.67 | |
</td> | </td> | ||
</tr> | </tr> | ||
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</td> | </td> | ||
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| - | + | 46.0 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.89 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.62 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 55.8 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.88 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.44 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 55.3 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.85 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.49 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 87.1 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.81 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.37 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 57.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.84 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.44 | |
</td> | </td> | ||
</tr> | </tr> | ||
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| - | + | 21.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.06 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.20 | |
</td> | </td> | ||
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| - | + | 16.2 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.13 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.20 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 47.4 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.78 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.16 | |
</td> | </td> | ||
</tr> | </tr> | ||
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</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 63.2 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.78 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.16 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 25.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.02 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.29 | |
</td> | </td> | ||
</tr> | </tr> | ||
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<td align="center"> | <td align="center"> | ||
| - | + | 27.0 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 2.09 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.78 | |
</td> | </td> | ||
</tr> | </tr> | ||
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</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 79.7 ng/µL | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.86 | |
</td> | </td> | ||
<td align="center"> | <td align="center"> | ||
| - | + | 1.60 | |
</td> | </td> | ||
</tr> | </tr> | ||
Revision as of 17:18, 13 August 2013
Week 9: 12th August - 18th August
TECAN
Using the TECAN analysis, we are measuring the capacity of repression of the natural binding site that we extract from the E.coli's genome.
Preculture with M9 medium (with carbenicillin) have been launched for BL21 transformed with our 1st construction:
- Natural Fur Binding Site of Fec A + sfGFP (clone 1, 2, 3)
- Natural Fur Binding Site of Fep A + sfGFP (clone 1, 2, 3)
- Natural Fur Binding Site of Ace B + sfGFP (clone 1, 2, 3)
Note: Preculture have been made in M9 medium with iron in oder to inhibit the expression of sfGFP.
After one night of culture, time the precultures have been refreshed by diluting them 200 times in M9 medium (with iron and carbenicillin).
2nd construction
Sequencing preparation
Preculture with LB medium (with carbenicillin) have been launched for Top10 transformed with our 2nd construction, then minipreps have been realized using the miniprep kit from Machrey Nagel.
| Name | Clone | Concentration | 260/280 | 260/230 |
|---|---|---|---|---|
| Fur Binding Site n°1 | 1 | 50.7 ng/µL | 1.62 | 0.68 |
| 2 | 21.1 ng/µL | 1.69 | 0.44 | |
| 3 | 21.6 ng/µL | 1.20 | 0.13 | |
| 4 | 70.8 ng/µL | 1.66 | 0.72 | |
| Fur Binding Site n°2 | 1 | 39.1 ng/µL | 1.23 | 0.40 |
| 2 | 25.1 ng/µL | 1.37 | 0.29 | |
| 3 | 61.1 ng/µL | 1.57 | 0.71 | |
| 4 | 54.7 ng/µL | 1.62 | 0.68 | |
| Fur Binding Site n°3 | 2 | 38.4 ng/µL | 1.84 | 0.78 |
| 3 | 114.6 ng/µL | 1.66 | 0.78 | |
| 4 | 39.2 ng/µL | 1.87 | 1.21 | |
| Fur Binding Site n°5 | 1 | 77.7 ng/µL | 1.76 | 0.99 |
| 3 | 78.4 ng/µL | 1.61 | 0.81 | |
| 4 | 72.7 | 1.85 | 4.23 | |
| Fur Binding Site n°6 | 1 | 70.4 ng/µL | 1.76 | 2.16 |
| 2 | 91.5 ng/µL | 1.55 | 1.21 | |
| 3 | 65.4 ng/µL | 1.70 | 1.52 | |
| 4 | 85.6 ng/µL | 1.72 | 1.65 | |
| Fur Binding Site n°7 | 1 | 53.5 ng/µL | 1.76 | 1.93 |
| 2 | 77.6 ng/µL | 1.69 | 1.22 | |
| 3 | 72.3 ng/µL | 1.68 | 1.22 | |
| Fur Binding Site n°8 | 1 | 73.9 ng/µL | 1.58 | 0.84 |
| 2 | 51.3 ng/µL | 1.75 | 1.10 | |
| 3 | 45.9 ng/µL | 1.85 | 1.43 | |
| 4 | 46.1 ng/µL | 1.88 | 1.61 | |
| Fur Binding Site n°9 | 1 | 108.3 ng/µL | 1.78 | 1.37 |
| 2 | 67.0 ng/µL | 1.88 | 1.61 | |
| 3 | 103.5 ng/µL | 1.87 | 1.79 | |
| 4 | 28.2 ng/µL | 2.06 | 1.76 | |
| Fur Binding Site n°10 | 1 | 29.5 ng/µL | 1.95 | 1.22 |
| 2 | 70.0 ng/µL | 1.90 | 1.67 | |
| 3 | 46.0 ng/µL | 1.89 | 1.62 | |
| 4 | 55.8 ng/µL | 1.88 | 1.44 | |
| Fur Binding Site n°11 | 1 | 55.3 ng/µL | 1.85 | 1.49 |
| 2 | 87.1 ng/µL | 1.81 | 1.37 | |
| 3 | 57.7 ng/µL | 1.84 | 1.44 | |
| 4 | 21.7 ng/µL | 2.06 | 1.20 | |
| Fur Binding Site n°15 | 1 | 16.2 ng/µL | 2.13 | 1.20 |
| 3 | 47.4 ng/µL | 1.78 | 1.16 | |
| 4 | 63.2 ng/µL | 1.78 | 1.16 |
Note: Fur Binding Sites n°4, 12, 13 and 14 are missing.
3rd construction
Sequencing preparation
Preculture with LB medium (with carbenicillin) have been launched for Top10 transformed with our plasmid which contains the sfGFP under the control of Lac O promoter. After one night of culture minipreps of 3 different clones have been realized using the miniprep kit from Machrey Nagel.
| Clone | Concentration | 260/280 | 260/230 |
|---|---|---|---|
| 1 | 25.7 ng/µL | 2.02 | 2.29 |
| 2 | 27.0 ng/µL | 2.09 | 1.78 |
| 3 | 79.7 ng/µL | 1.86 | 1.60 |
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