What reaction would you expect when performing a positive control in the oxidase assay? What would it mean if a known oxidase-positive bacterium did not cause the expected reaction?
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- What reaction would you expect when performing a positive control in the oxidase assay? What would it mean if a known oxidase-positive bacterium did not cause the expected reaction?
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- What reaction would you expect when performing a positive control in the catalase assay? What would it mean if a known catalase-positive bacteria did not produce the expected reaction?Why is it essential to add sodium azide to succinate dehydrogenase assay solution? What is the purpose of including the sodium succinate solution?Using a schematic diagram, summarize the following steps in preparing competent cells for transformation: Inoculate a single colony of E. coli into 5 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm). Add 200 μl of the culture into 50 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm) to an OD600 = 1.3 to 1.5. Aliquot culture into five 15-ml pre-chilled, conical tubes. Leave tube on ice 5 to 10 min. Centrifuge cells 7 min at 1,600 × g (3,000 rpm), 4°C. Pour off supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution (50 mM CaCl2), perform resuspension very gently, and keep on ice. Centrifuge cells 5 min at 1,100 × g (2,500 rpm), 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Keep resuspended on ice for 30 min. Centrifuge cells 5 min at 1,100 × g, 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Dispense cells (250 μl) into pre-chilled, sterile…
- For the study of alanine production by a recombinant strain of E. coli, cultivation was carried out in a benchtop bioreactor with 4.5 L of culture medium, using glucose as a limiting substrate. During the cultivation, there was no lag phase and the cells showed exponential growth for 5 hours. The following table presents the results of the analysis of ammonia and glucose consumption, and alanine accumulation throughout the cultivation. Knowing that 500 mL of a cell suspension at a concentration of 5.0 g/L (inoculum) was added to the 4.5 L of medium in the reactor and that the YX/NH3 previously determined was 7.5, calculate:a) the maximum specific growth rateb) YX/S and YP/S yield factorsc) How long would it take to reach Cx = 30 g/L if the cells continued with the exponential growth profile until the end of the culture (without nutrient deprivation or any type of inhibition)?d) Describe how the mathematical treatment of the data should be done to determine the type of product formation…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 II 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.Your colleague handed you a novel strain of coli that is purifying a protein with a 6xHisTag; they claim it is superior to the TOP10 cells you have been using. But even with a larger culture size, you discover that your protein yield—using the same Ni-NTA column—is quite low. You discover that the novel strain of Escherichia coli generates an unusually high quantity of dicarboxylic acids, a byproduct of the citric acid cycle that is recognized for its ability to function as an all-purpose metal chelator. What do you think the issue is with purifying IMAC protein with this new strain of E. coli?
- What is the purpose of the negative and positive control in the Disc diffusion assay? Why is it important to use positive and negative controls in microbial experimentation? What does it tell us if the positive control leaves a zone of inhibition but the experimental treatment (penicillin disc) does not?Why do we need to determine the extinction coefficient in order to calculate the initial velocity in an enzyme kinteic assay experiment?In the Avery, McLeod, McCarty Experiment where supernatant from heat killed, virulent S Strain pneumonia solutions were added to non-virulent R Strain pneumonia cell cultures and allowed to grow in liquid media (i.e., broth). In tubes where Protease was added to the supernatant prior to cell culture, what was the observed effect when plating and growing the S. pneumonia cells to solid media?
- In the Avery, McLeod, McCarty Experiment where supernatant from heat killed, virulent S Strain pneumonia solutions were added to non-virulent R Strain pneumonia cell cultures and allowed to grow in liquid media (i.e., broth). In tubes where Protease was added to the supernatant prior to cell culture, what was the observed effect when plating and growing the S. pneumonia cells to solid media? Selected answer will be automatically saved. For keyboard navigation, press up/down arrow keys to select an answer. a b C d e All RNA was degraded and Transformation of the R Strain to S Strain occurred. All Protein was degraded and Transformation of the R Strain to S Strain occurred. All DNA was degraded and Transformation of the R Strain to S Strain occurred. All RNA was degraded and no Transformation occurred indicating RNA is the molecule of Transformation inheritance None of the above are trueDescribe the p-nitrophenol assay and how a standard curve can be used to calculate the concentration of an unknown .You got an opportunity to join a professor lab who is working in-vivo model and specifically looking at the dysregulation of mitochondria in liver. He asked you to isolate mitochondria from a Rat liver and placed in an assay medium. Based on the knowledge you gain in this course so far, please answer the following questions: a) Which technique will you use to isolate mitochondria? b) What happens to the pH of the medium when the medium is kept anaerobic? c) What happens when O2-saturated saline is added to the mixture?