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Team:Calgary/Notebook/Protocols/decatecholization
From 2012.igem.org
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<p>This assay is used to verify that catechol 2,3-dioxygenase (XylE) is converting catechol to the yellow compound 2-hydroxymuconic semialdehyde (2-HMS). For this procedure we used to the newly constructed XylE part. This part contains the TetR promoter the XylE gene and its native rbs site:</p> | <p>This assay is used to verify that catechol 2,3-dioxygenase (XylE) is converting catechol to the yellow compound 2-hydroxymuconic semialdehyde (2-HMS). For this procedure we used to the newly constructed XylE part. This part contains the TetR promoter the XylE gene and its native rbs site:</p> | ||
<ol> | <ol> | ||
| - | <li>Grow up 2 ml overnight cultures in LB</li> | + | <li>Grow up 2 ml overnight cultures in LB.</li> |
| - | <li>Spin the cultures down and keep the supernatant </li> | + | <li>Spin the cultures down and keep the supernatant.</li> |
| - | <li>Bring the supernatant to a concentration of 0.1 M of Catechol by using a 1M catechol stock solution</li> | + | <li>Bring the supernatant to a concentration of 0.1 M of Catechol by using a 1M catechol stock solution.</li> |
| - | <li>The colour of the supernatant should change to bright yellow very quickly ( about 30 seconds) | + | <li>The colour of the supernatant should change to bright yellow very quickly (in about 30 seconds).</li> |
</ol> | </ol> | ||
<br><br><br><br><br><br><br><br><br> | <br><br><br><br><br><br><br><br><br> | ||
</html>}} | </html>}} | ||
Revision as of 02:10, 2 October 2012
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Catechol Assay E. coli Cells
This assay is used to verify that catechol 2,3-dioxygenase (XylE) is converting catechol to the yellow compound 2-hydroxymuconic semialdehyde (2-HMS). For this procedure we used to the newly constructed XylE part. This part contains the TetR promoter the XylE gene and its native rbs site:
- Grow up 2 ml overnight cultures in LB.
- Spin the cultures down and keep the supernatant.
- Bring the supernatant to a concentration of 0.1 M of Catechol by using a 1M catechol stock solution.
- The colour of the supernatant should change to bright yellow very quickly (in about 30 seconds).
