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- 14. What is the null and alternative hypothesis of Bradford protein assay?1. Describe the basics of flow cytometry. How is this technology used to sort cells? 2. Describe an experiment demonstrating the utility of cell sorting in biotechnology. 3. How do you think flow cytometry can be used as a medical application? What are some examples where flow cytometry is used in the treatment or diagnosis of specific diseases?1. You are studying a biochemical pathway and isolate Neurospora mutants I, II, and III. Mutant I can grow if you supplement the medium with Z. Mutant Il can grow if you supplement the medium with X, Y, or Z. Mutant III can grow if you supplement the medium with X and Z, but not with Y. Draw a biochemical pathway that shows the correct order for compounds X, Y, and Z and for the enzymes that each mutant is defective for.
- 12. When a person is determining the concentration of a sample, they will frequently repeat the samemeasurement of a sample by preparing three or more identical tubes (i.e. make replicates).a) Although you did not set up replicates today, what would be the purpose of setting upreplicates?b) If you had tested three replicates of test tube #2 in the determination of glucoseconcentration, what would you do with the data from the three replicates?Match description with Term: 1) determine the amount of protein in a sample 2) determine the location of a protein in a cell or sample 3) determine the amount of transcript in a sample 4) Determine the location of a transcript in a sample or cell a. In situ hybridization b. northern blotting c. western blotting d. immunocytochemistry4) Researcher #1 and #2 in your lab each conducted a protein purification. They present you with the following data. The protein-of-interest is indicated by the red arrowhead, and the numbers on top indicate the purification step. Researcher #1 1 2 3 4 SDS-PAGE (Coomassie Stained) Researcher #2 1 2 3 4 5 6 Immunoblot a) Which researcher has more clearly demonstrated purification of the protein-of-interest? Justify your answer. b) What type of chromatography step did researcher #1 most likely conduct in going from step 2➜ 3 based on the gel? Justify your answer.
- What is a CBC? In your own words, briefly describe why it is useful.4) Consider the following list of hypothetical antibiotics, tested against Escherichia coli in a Kirby-Bauer test: Antibiotic Zone of Inhibition (mm) Astonostatin 12 Bodaciosporin 25 Crazifloxacin 5 Dorkimycin 16 When the above antibiotics were tested as treatments for E. coli infections in humans, it was found that Dorkimycin has virtually no effect on the infection, while Astonostatin is the most effective of the four antibiotics listed. Propose an explanation for this result. Based purely on these results, can you draw up a similar table for predicted zones f inhibition for Staphylococcus aureus? Why or why not?4) List 3 things that a potential vector must have in order to be useful. 5) Sometimes the gene for the enzyme glutathione-S-transferase (GST) is used in purifying your protein of interest. Briefly explain how this is done. 6) What enzyme is used to create CDNA from MRNA?
- Which of the following statements is/are FALSE about Bradford assay? 1. Upon addition of the reagent, the color of the protein solution becomes purple. II. The absorbance of the resulting solution is measured at a wavelength of 540-550 nm. Both I and II O II only Neither I nor II O I onlyQ.No.4. Describe the critical steps required to make a recombinant bacterial cell for production of commercial products.(subject: Genetic engineering).4. The image below is based on the Meselson Stahl experiment where bacteria are grown in 15N and moved to 14N. Please match the tube in the image to the descriptors a-e a) DNA from E. coli grown in 15N test tube 1 test tube 2 test tube 3 test tube 4 test tube 5 b) DNA from E. coli grown in 14N c) A 1:1 mixture of DNA from cells grown in 14N and cells grown in 15N d) A 1:1 mixture of DNA grown in 14N and 15N, heated and allowed to re- anneal e) DNA containing one strand of 15N and one strand of 14N