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Team:Warsaw/Calendar-Main/20 July 2009
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(New page: {{WarNotebook}} <!-- do not edit above me! --> <html> <h3>Insertion of the pho gene into the pKSII+ plasmid</h3> <h4>Kama</h4> <li>Plasmid digest mix was prepared as fallows: 3μl Tang...) |
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<li>Pho gene digest mix was prepared as fallows: 3μl Tango buffer (Fermentas), 16μl purified gene, 1μl XbaI enzyme, the solution was topped up with H2O to the final volume of 30 μl.</li> | <li>Pho gene digest mix was prepared as fallows: 3μl Tango buffer (Fermentas), 16μl purified gene, 1μl XbaI enzyme, the solution was topped up with H2O to the final volume of 30 μl.</li> | ||
<li>The digest was kept for 2h at 30°C, after that 2h at 37°C and then the enzymes were inactivated for 20min. at 70°C.</li> | <li>The digest was kept for 2h at 30°C, after that 2h at 37°C and then the enzymes were inactivated for 20min. at 70°C.</li> | ||
| - | + | <li><p>The ligation mix was prepared as follows: 2μl plasmid, 8μl gene, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. As a control:2μl plasmid, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. The ligation was carried out in 18°C overnight (~15h)</li> | |
</html> | </html> | ||
Revision as of 22:32, 21 July 2009
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Insertion of the pho gene into the pKSII+ plasmid
Kama
The ligation mix was prepared as follows: 2μl plasmid, 8μl gene, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. As a control:2μl plasmid, 3μl ligation buffer, 1 μl T4 DNA ligase (Fermentas), 0,5μl PEG the solution was topped up with H2O to the final volume of 30 μl. The ligation was carried out in 18°C overnight (~15h)
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