BIOC 301 Final Exam Mock Questions.docx

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BIOC 301
Jason Read

Final Exam Mock Questions 1 After 3 years of graduate studies, you have managed to purify a protein to the point where you are able to crystalise it. At this point, you can assume that the protein is, essentially, pure. You decide to work out how much of the protein you actually have. Using an online tool (which is not down for website maintenance...) you determine that the expected extinction co-efficient at 280nm for the protein under your spectrophotometry conditions is 250 00 M -1 , cm -1 . You prepare 100ul of a 1:10 dilution of your protein and do a spec reading at 280nm; you get a value of 0.153. Given that you have 1ml of the undiluted protein solution and that the protein has a molecular mass of 150kDa how many micrograms of protein have you wasted spent 3 years isolating? 2 Given the following graph, the formula in the enzyme kinetics documents and that the inhibitor concentration was 4mM, determine Km and Ki for this system (substrate, enzyme, inhibitor). 3 Given the following information and the following enzymes, what enzymes could you use to perform a restriction digest? What would you expect the sizes of fragments to be? Assume complete digestion of plasmids. Assume plasmid was ligated into the plasmid using the HindII site. Assume all restriction digest sites are at 425 for ease of calculation. 4 NaCl costs $50 for 100 grams. Na2HPO4 costs $100 for 500 grams. CH3COOK costs $75 for 200 grams. Assuming the water is free, how much does 750ml of the following solution cost? Buffer X= 1.5M NaCl, 0.5M Na2HPO4, 100mM CH3COOK 5 Given the following absorbance values for the ONPG Assay (and different substrate concentration and over time) and the Bradford Assay for an unknown enzyme D, answer the following questions. ONPG 405 nm Time Absorbance 2 0.524 4 0.849 6 1.03 8 1.358 10 1.6 12 1.743 14 1.854 16 1.949 ONPG 405 nm Dilution Absorbance 1 1.419 2 0.795 4 0.476 8 0.209 16 0.124 32 0.066 Bradford 595 nm Concentration (ug) Absorbance 0 0.512 1 0.636 3 0.674 5 0.77 10 0.947 a Construct a Michaelis-Menten Curve. Given that the original substrate concentration was 10 mM. b Construct a Lineweaver-Burk Curve. What is the initial velocity of the reaction? What is the Vmax? What is the Km of the substance? Does the reaction rate stay constant throughout the experiment? c Construct the standard curve for the Bradford Assay. Based on the curve, what is the protein concentration? (Absorbance of sample = 0.578, 10 ul of a 1:15 dilution) d What is the specific activity of the reaction? 6 Given the following pre and
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