Team:Calgary/Notebook/Protocols/decatecholization

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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 ( about 30 seconds)s

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