Lab 10: MINI LAB REPORT
Refer to the general guidelines for Mini Lab Reports on Canvas, found here (link opens in a new tab): Mini Lab Reports
Methods
As always, describe the assays in enough detail that someone with scientific training could repeat it. Include the assay conditions and what you were assaying. Describe how the assay was set up including a description of the blank. State the concentrations of PNPA you used. State how many replicates were done.
Note that you do not need to explicitly describe the pilot experiments. Generally methods sections don’t include details on how an assay was optimized, or any troubleshooting that may have been done. Simply report what was actually done to generate the data you are showing in the report.
State how you calculated all the values you need to report in the results section and be sure to include equations.
Results
- Plot of V0 vs. [S] from Prism. Be sure to label your axes.
- In the text, state the results of the fit including the Vmax and KM. Include error measurements.
- Table of relevant values, including kcat and kcat/ KM
- Always remember to include error measurements and p-values where applicable.
Discussion
- Discuss the kcat/KM values – what do they tell you about the enzyme?
- Compare the KM, kcat and kcat/KM of the wild type and mutant HCAII. Discuss how the mutation may be affecting the activity of the protein. Be specific. Include information about the mutated amino acid in your answer. Was the result expected based upon the structural and biochemical properties of the mutation? Why or why not?
- How could you improve this experiment? For this discussion point, consider the concentration range of the substrate, the concentration of wild type and mutant HCAII, the HCAII mutant you assayed, the order in which you assembled your samples, etc.