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Team:Warsaw/Calendar-Main/30 April 2009
From 2009.igem.org
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| + | <h3>Digestion of the pKS/hly contructs</h3> | ||
| + | <h4><font color="black">Kamil</font></h4> | ||
| + | <br /> | ||
| + | <p>Tasks:</p> | ||
| + | <ul> | ||
| + | <li>Verify the correctness of the pKS/hly constructs</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | <p>Methods:</p> | ||
| + | <ul> | ||
| + | <li> The BanI enzyme was selected for the test because the hly insert bears an additional digestion site.</li> | ||
| + | <li><p>Digestion mixture's composition:</p> 2ul Orange buffer (Fermentas), 1ul BanI enzyme, 1ul plasmid | ||
| + | |||
| + | solution,solution was topped up with H2O to 20ul. </li> | ||
| + | <li> The digestion was kept at 37 °C overnight (~18h) and then inactivated at 65 °C for 10 min.</li> | ||
| + | <li>The electrophoretic separation was carried out on 1% agarose gel</li> | ||
| + | </ul> | ||
| + | <br /> | ||
| + | <p>Results:</p> | ||
| + | <aling="centre"><img src="https://static.igem.org/mediawiki/2009/6/66/2009.04.30_-_trawienie_konstruktow_hly_opisany.jpg"/> | ||
| + | |||
| + | |||
| + | </aling> | ||
| + | <var> | ||
| + | <ul> | ||
| + | <li>Gel (from left)</li> | ||
| + | </ul> | ||
| + | <ol> | ||
| + | <li>GeneRuler DNA Ladder Mix #SM0333 (Fermentas) (3ul)</li> | ||
| + | <li>sample no. 15 (5ul)</li> | ||
| + | <li>sample no. 16 (5ul)</li> | ||
| + | <li>sample no. 17 (5ul)</li> | ||
| + | <li>sample no. 18 (5ul)</li> | ||
| + | <li>pKS plasmid (control) (5ul)</li> | ||
| + | |||
| + | |||
| + | </ol> | ||
| + | </var> | ||
| + | <br /> | ||
| + | <p>Notes:</p> | ||
| + | <ul> | ||
| + | <li>At least 5ul of plasmid solution should have been used</li> | ||
| + | </ul> | ||
| + | <br /> | ||
</html> | </html> | ||
Revision as of 20:10, 18 June 2009
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Digestion of the pKS/hly contructs
Kamil
Tasks:
- Verify the correctness of the pKS/hly constructs
Methods:
- The BanI enzyme was selected for the test because the hly insert bears an additional digestion site.
Digestion mixture's composition:
2ul Orange buffer (Fermentas), 1ul BanI enzyme, 1ul plasmid solution,solution was topped up with H2O to 20ul.- The digestion was kept at 37 °C overnight (~18h) and then inactivated at 65 °C for 10 min.
- The electrophoretic separation was carried out on 1% agarose gel
Results:
- Gel (from left)
- GeneRuler DNA Ladder Mix #SM0333 (Fermentas) (3ul)
- sample no. 15 (5ul)
- sample no. 16 (5ul)
- sample no. 17 (5ul)
- sample no. 18 (5ul)
- pKS plasmid (control) (5ul)
Notes:
- At least 5ul of plasmid solution should have been used
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