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Team:Paris Liliane Bettencourt/Notebook/2010/07/06/
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(New page: {{Template:paris2010_2}} <p style="display:block;"><font color="red">Raphaël, Théotime and Stéphane</font> </p><p style="display:block;"><u>Miniprep - Kit Qiagen - Final volume :...) |
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</li><li>10 µL of ladder 1Kb | </li><li>10 µL of ladder 1Kb | ||
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| - | <p style="display:block;">Migration : 50V, 1h30 | + | <p style="display:block;">Migration : 50V, 1h30<br /> |
| - | </p><p style="display:block;"> | + | The bands of the ladder are not well separated. Thus, the migration should last longer and the gel should be at 1,0% agarose. |
| - | <br /> | + | </p><p style="display:block;">https://static.igem.org/mediawiki/2010/9/99/Electrophoresis_060710.jpg |
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Latest revision as of 19:26, 26 October 2010
Raphaël, Théotime and Stéphane
Miniprep - Kit Qiagen - Final volume : 50µL (Raphaël)
- pSUlib - attC KanR
Miniprep - Kit Promega (Stéphane)
- pSUlib - attC KanR
Restriction digest - Xba I
- pSUlib - attC KanR (Qiagen and Promega Minipreps)
Final volume : 20 µL
15 µL distilled water
2 µL minipreped DNA
0,2 µL BSA 100x
2 µL buffer 10x
1 µL restriction enzyme
Protocol
1- Add in order listed above.
2- Vortex the mix 2 sec, spindown in micro-centrifuge and put in 37°C for 2h.
DNA Gel electrophoresis (agarose 1,5% w/v)
7,5g of agarose in 500mL of TBE buffer 0,5x
2 µL of EtBr was added in the gel
Protocol (2 wells)
- 10 µL of minipreped DNA (Qiagen) + 2 µL of loading buffer 6x
- 10 µL of ladder 1Kb
Migration : 50V, 1h30
DNA Gel electrophoresis (agarose 1,0% w/v)
2 µL of EtBr was added in the gel
Protocol (2 wells)
- 10 µL of minipreped DNA (Promega) + 2 µL of loading buffer 6x
- 10 µL of ladder 1Kb
Migration : 50V, 1h30
The bands of the ladder are not well separated. Thus, the migration should last longer and the gel should be at 1,0% agarose.
