Team:Calgary/Notebook/Protocols/decatecholization

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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>
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<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)s </li>
+
<li>The colour of the supernatant should change to  bright yellow very quickly (in about 30 seconds).</li>
</ol>
</ol>
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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:

  1. Grow up 2 ml overnight cultures in LB.
  2. Spin the cultures down and keep the supernatant.
  3. Bring the supernatant to a concentration of 0.1 M of Catechol by using a 1M catechol stock solution.
  4. The colour of the supernatant should change to bright yellow very quickly (in about 30 seconds).