Team:Lethbridge/Notebook/Lab Work/September
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<b>Objective:</b> Characterized catechol by xylE enzyme<br> | <b>Objective:</b> Characterized catechol by xylE enzyme<br> | ||
- | <b>Method:</b> Measured absorbance of catechol (275nm) and 2-hydroxymuconate semialdehyde (380nm). | + | <b>Method:</b> Measured absorbance of catechol (275nm) and 2-hydroxymuconate semialdehyde (380nm).<br> |
- | + | *<b>Protocol:</b> | |
- | + | *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 50µM;). | |
+ | *4) Quench the reaction with 5A% w/v trichloroacetate at certain time points. (0,15sec, 30sec, 45sec, 60sec, 2min). | ||
+ | *5) Spin down cells. | ||
+ | *6) Measure absorbance of supernatant. |