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Team:Warsaw/Calendar-Main/24 July 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>Cloning of the mgtc promoter into the pKSII+ plasmid</h3> <h4>Kamil</h4> <br /> <p>Tasks:</p> <ul> <li>Plasmid purification</li> ...) |
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<li>Despite the fact that all transferred colonies turned more or less blue the purification was carried out using the Plasmid Mini kit (A&A Biotechnology) according to the manufacturers protocol.</li> | <li>Despite the fact that all transferred colonies turned more or less blue the purification was carried out using the Plasmid Mini kit (A&A Biotechnology) according to the manufacturers protocol.</li> | ||
<li>The digest mix was prepared as fallows: 10μl of purified plasmid, 2μl Tango buffer (Fermentas), 1μl SpeI enzyme, 1μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl.</li> | <li>The digest mix was prepared as fallows: 10μl of purified plasmid, 2μl Tango buffer (Fermentas), 1μl SpeI enzyme, 1μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl.</li> | ||
| + | <li>The digest was incubated for 3h in 37°C. | ||
| + | <li>The results were visualised with gel electrophoresis on 1% agarose gel. | ||
</ul> | </ul> | ||
<br /> | <br /> | ||
<p>Results:</p> | <p>Results:</p> | ||
<ul> | <ul> | ||
| - | <li> | + | <li>Gel Electrophoresis:</li> |
</ul> | </ul> | ||
<br /> | <br /> | ||
Revision as of 12:28, 24 July 2009
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Cloning of the mgtc promoter into the pKSII+ plasmid
Kamil
Tasks:
- Plasmid purification
- Control digest
Methods:
- Despite the fact that all transferred colonies turned more or less blue the purification was carried out using the Plasmid Mini kit (A&A Biotechnology) according to the manufacturers protocol.
- The digest mix was prepared as fallows: 10μl of purified plasmid, 2μl Tango buffer (Fermentas), 1μl SpeI enzyme, 1μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl.
- The digest was incubated for 3h in 37°C.
- The results were visualised with gel electrophoresis on 1% agarose gel.
Results:
- Gel Electrophoresis:
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