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Team:Calgary/Notebook/Protocols/decatecholization
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Revision as of 04:53, 3 October 2012
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Catechol Assay in 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).
