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(Solved by Expert Tutors) Hi. I need to make Michaelis-Menten plot and I need help to do it.


I performed spectrophotometry on 6 cuvettes containing NAD+, Buffer, Ethanol, H2O, and ADH, with varying concentrations of ethanol.

I have the slopes of the 6 graphs I made from these UV specs. Now I need help turning these slopes into Vo values. In order to make the Michaelis-Menten graphs, I need to plot Vo values vs. [Ethanol]. I believe this will involve the Beer's equation, but I'm not sure how to proceed and I need help from someone with a good understanding of Michaelis-Menton plots. The wavelength used was 340nm, and the E340nm for NADH = 6220 M-1cm-1.

My slopes are as follows:

Cuvette 1: 0.004

Cuvette 2: 0.006

Cuvette 3: 0.008

Cuvette 4: 0.009

Cuvette 5: 0.001 *outlier

Cuvette 6: 0.010

The ethanol concentrations were as follows:

cuvette 1: 25 microliters

cuvette 2: 50 microliters

cuvette 3: 125 microliters

cuvette 4: 250 microliters

cuvette 5: 375 microliters

cuvette 6: 750 microliters

If you can help me to find the Vo values for my plot that would be great. Please let me know if I can provide anymore helpful information. Thank you.

 


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Apr 19, 2020

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