This exercise is to let you go through the same analysis processes originally designed for Experiment 3. Initially each student group will purify and characterize one G6PD variant.
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Description
This exercise is to let you go through the same analysis processes originally designed for Experiment 3. Initially each student group will purify and characterize one G6PD variant. It involves purification of his-tagged protein, determination of enzyme activity, protein assay and calculation of enzyme specific activity. Then the specific enzyme activity data from various groups are combined to see whether there is any significant difference in the enzyme activity among the three G6PD variants.
The exercise is based on the procedures described in practical manual on the purification and characterization of his-tagged human G6PD expressed in E. coli cells.
Please refer to page 3 to page 7 of the practical manual for details on the procedure and calculation. The data below were the actual raw data reported by one
student group before (so there may be some flaws in the data). Note that the volume of samples and the dilution factor used may differ from that listed in the
manual. You will need to adjust the formula accordingly when calculating the enzyme activity and protein concentration.
