Team:Lethbridge/Notebook/Lab Work/September

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(→September 23, 2010)
(→September 23, 2010)
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*2) Take 1/10 dilution of cells
*2) Take 1/10 dilution of cells
*3) Introduce 1µL of 0.05M catechol solution into the cell dilution. (Final concentration of 50&microM;).
*3) Introduce 1µL of 0.05M catechol solution into the cell dilution. (Final concentration of 50&microM;).
-
*4) Quench the reaction with 5A% w/v trichloroacetate at certain time points. (0,15sec, 30sec, 45sec, 60sec, 2min).
+
*4) Quench the reaction with 5A% w/v trichloroacetate at certain time points. (0,15sec, 30sec, 45sec, 60sec, 2min, 3min, 4min, 5min, 10min).
*5) Spin down cells.
*5) Spin down cells.
*6) Measure absorbance of supernatant.<br>
*6) Measure absorbance of supernatant.<br>
<b>Results:</b>
<b>Results:</b>

Revision as of 04:06, 24 September 2010



Feel free to look around our notebook!

Here you can check out the work we have done in the lab, click on a month to take a look!

September 23, 2010

( JV)

Objective: Characterized catechol degradation by xylE enzyme

Method: Measured absorbance of catechol (275nm) and 2-hydroxymuconate semialdehyde (380nm).

  • Protocol:
  • 1) Grow cells in M9 minimal medium
  • 2) Take 1/10 dilution of cells
  • 3) Introduce 1µL of 0.05M catechol solution into the cell dilution. (Final concentration of 50&microM;).
  • 4) Quench the reaction with 5A% w/v trichloroacetate at certain time points. (0,15sec, 30sec, 45sec, 60sec, 2min, 3min, 4min, 5min, 10min).
  • 5) Spin down cells.
  • 6) Measure absorbance of supernatant.

Results:

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