Biochem Lab Exam 1 Review
Lab 1 (Lab Calculations)
1. Be able to use the concept of ratios and proportions to solve a common laboratory calculation.
2. Be able to use the concept of percents to solve a common laboratory calculation.
3. Be able to use the concept of dilutions to solve a common laboratory procedure like single dilutions,
multi-component dilutions, independent dilution series (step dilutions), or serial dilutions.
4. Be able to solve common laboratory calculations involving concentrations that are expressed as %
weight per volume, % volume per volume, weight per volume, molarity, and “X”.
5. Be able to accurately make a molar solution from a dry chemical stock.
6. Be able to accurately make a dilution of a stock solution using either a pipetman or a pipet aid.
7. Be able to accurately make a multi-component solution using either a pipetman or a pipet aid.
8. Be able to accurately perform a serial dilution using either a pipetman or a pipet aid.
9. Know and apply the common concepts to avoid error while using pipetman and laboratory balances.
10. Given a variety of common laboratory glassware, utilize the most accurate when preparing solutions.
11. Know and be able to apply the differences between the terms: accuracy, precision, standard
deviation, and reproducibility.
12. Know graphically and statistically what standard deviation means and how it relates to precision and
reproducibility.
13. Be able to calculate the average the standard deviation for a set of data.
Lab 2 (Alfalfa Sprout Phosphatase Activity)
1. What is the physiological role of phosphatases?
2. What are the two main groups of phosphatases and how are they classified?
3. What is the basis of the reaction to detect the presence of phosphatase activity?
4. What is the purpose of the sodium carbonate?
5. Be able to interpret graphs that depict enzyme activity with respect to changes in pH, temperature,
and reaction time.
6. Why is it important to consider the application of a purified protein prior to actually designing a
purification scheme?
7. What are the four intellectual steps involved in optimizing the purification of a protein?
8. Why should you “never follow a seal act with a seal act”? (hint… this concept is in your background
readings)
9. When might you need to use a coupled reaction?
10. When might you need to use a continuous time course assay?
11. Be able to design an enzyme assay based on the reaction the enzyme catalyzed.
Lab 3 (Expression rGFP in E. coli)
1. What is GFP? How/why does GFP “glow” green?
2. Why was the GFPUV gene placed into the pRSETA plasmid?
3. How is a “calculated MW” and an “estimated MW” of a protein determined?
4. Why was the pRSETA-GFPUV transformed into the E. coli strain BL21<DE3>pLysS?
5. What is the general process of inducing expression of rGFP?
6. Why was the GFP placed under the control of a T7 promoter?
7. What is the purpose of adding IPTG to the growing culture?
8. What was the purpose of the pLysS plasmid?
9. What is the purpose of adding ampicillin and chloramphenicol to the media?
10. Why do we take culture samples before and after the addition of IPTG?
11. Who first discovered GFP?
12. Who first cloned GFP?
13. What was the first transgenic organism to express GFP?
14. What was the first transgenic eukaryote to express GFP?