Difference between revisions of "Team:Tuebingen/Experiments"
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<ul> | <ul> | ||
<li>Following electrophoresis, excise DNA band from gel and place gel slice in a 1,5ml microcentrifuge tube.</li> | <li>Following electrophoresis, excise DNA band from gel and place gel slice in a 1,5ml microcentrifuge tube.</li> | ||
− | <li>Add 10μ</li> | + | <li>Add 10μl Membrane Binding Solution per 10mg of gel slice. Vortex and incubate at 50-60°C until gel slice is completely dissolved.</li> |
− | <li></li> | + | <li>Insert SV Minicolumn into Collection Tube.</li> |
− | <li></li> | + | <li>Transfer dissolved gel mixture to the Minicolumn assembly. Incubate at room temp. for 1 minute.</li> |
− | <li></li> | + | <li>Centrifuge at 16,000xg for 1 min. Discard flowthrough and reinsert Minicolumn into Collection tube.</li> |
− | <li></li> | + | <li>Add 700μl Membrane Wash Solution (ethanol added). Centrifuge at 16,000xg for 1 min. Discard flowthrough and reinsert Minicolumn into Collection tube. </li> |
− | <li></li> | + | <li>Repeat this step with 500μl Membrane Wash Solution. Centrifuge at 16,000xg for 5 min.</li> |
− | <li></li> | + | <li>Empty the Collection Tube and recentrifuge the column assembly for 1,5 min.</li> |
+ | <li>Leave the tubes open for 10 min (to let any rest of ethanol evaporate).</li> | ||
+ | <li>Carefully transfer Minicolumn to a clean 1.5ml microcentrifuge tube.</li> | ||
+ | <li>Add 30μl of Nuclease-Free-Water (65°C) to the Minicolumn. Incubate at 65°C for 5 min. Centrifuge at 16,000xg for 1 min.</li> | ||
+ | <li>Discard Minicolumn and store DNA at 4°C or -20°C.</li> | ||
</ul> | </ul> | ||
</div> | </div> |
Revision as of 09:04, 25 August 2015
![](https://static.igem.org/mediawiki/2015/b/ba/Team_Tuebingen_menu_coli_mit_lightstrahl.png)
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Protocols
Prep
Colony-PCR
3A-Assembly
Restriction
Transformation
PCR
Ligation
Testrestriktion
Gel extraction
DNA Purification using Promega Wizard SV Gel and PCR Clean-Up System Kit
This is a modified protocol for the Wizard SV Gel and PCR Clean-Up System Kit from Promega
- Following electrophoresis, excise DNA band from gel and place gel slice in a 1,5ml microcentrifuge tube.
- Add 10μl Membrane Binding Solution per 10mg of gel slice. Vortex and incubate at 50-60°C until gel slice is completely dissolved.
- Insert SV Minicolumn into Collection Tube.
- Transfer dissolved gel mixture to the Minicolumn assembly. Incubate at room temp. for 1 minute.
- Centrifuge at 16,000xg for 1 min. Discard flowthrough and reinsert Minicolumn into Collection tube.
- Add 700μl Membrane Wash Solution (ethanol added). Centrifuge at 16,000xg for 1 min. Discard flowthrough and reinsert Minicolumn into Collection tube.
- Repeat this step with 500μl Membrane Wash Solution. Centrifuge at 16,000xg for 5 min.
- Empty the Collection Tube and recentrifuge the column assembly for 1,5 min.
- Leave the tubes open for 10 min (to let any rest of ethanol evaporate).
- Carefully transfer Minicolumn to a clean 1.5ml microcentrifuge tube.
- Add 30μl of Nuclease-Free-Water (65°C) to the Minicolumn. Incubate at 65°C for 5 min. Centrifuge at 16,000xg for 1 min.
- Discard Minicolumn and store DNA at 4°C or -20°C.
Experiments & Protocols
Describe the experiments, research and protocols you used in your iGEM project.
What should this page contain?
- Protocols
- Experiments
- Documentation of the development of your project