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The bacteria that you are growing to express biochemisfunase are resistant to ampicillin, so you keep ampicillin in your media to prevent contamination from other bacterial strains. You find a tube of ampicillin in the lab freezer that says “2000x AMP” on the lid, which indicates that it is 2000 times more concentrated than what you need (i.e., a concentration of 1x ampicillin in your media). How much of the 2000x ampicillin stock would you add to your 1.0-L culture to get a final concentration of 1x ampicillin?
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- G418 disulfate (FW 692.7) is an antibiotic used to select transfected mammalian cells in tissue culture. The standard working concentration for selection is 400 µg/mL of active ingredient in culture medium. The bottle states that it contains 71% active ingredient. You need to make a 100x stock in medium and adjust the pH before you apply it to cells. How much powder should be added to make 10 mL of stock solution? How much do you add to a 2 mL culture dish?A vial of Doxorubicin reads 0•5g per vial. Instructions say to reconstitute each 12mg with 2•5ml of NS. How many ml of NS will be needed to reconstitute the vial of the recommended concentration? please show workingIn a nutrient medium that lacks histidine, a thin layer of agar containing ~109 Salmonella typhimurium histidine auxotrophs (mutant cells that require histidine to survive) produces ~13 colonies over a two-day incubation period at 37 °C. How do these colonies arise in the absence of histidine? The experiment is repeated in the presence of 0.4 μg of 2-aminoanthracene. The number of colonies produced over two days exceeds 10,000.What does this indicate about 2-aminoanthracene? What can you surmise about its carcinogenicity?
- Given this, if you used 6g of vitamin Z powder to make 20 ml of solution, what is the % concentration of this solution? (I gave the image since I don't know if that info is needed to solve this question.)It also gives a follow-up, if you can help here too: You work in a lab as a summer student. One of your tasks is to make sure that there is enough cell culture medium containing antibiotics to grow bacteria. One day you realize that there is only 5 ml of 10% Antibiotic stock solution in the freezer. You decide to use it all to prepare the working culture medium with 0.01% antibiotic. In the lab there is plenty of growth medium without antibiotics. (Note: dilution in medium is like dilution in water). You remember the equation to make dilutions of stock solutions. You usually use this formula to calculate the required volume of a stock solution, but you realize it can apply here as well, even though the unknown is the final volume. So, you make that dilution. Given that each bacterial…Each liter of an intranasal antibiotic solution contains 160 mg of gentamicin. You have 2-mL gentamicin vials that contain 40 mg. How many vials will you need if you are to prepare three liters of the antibiotic solution?You overexpressed (asked bacteria to make a lot of) an 80 kDa protein carrying a 6X-His tag and purified it on a nickel column. You collect frac ons from your purifica on and analyze them using SDSPAGE. The results are shown in image attached. a) Interpret the results from the gel. What is wrong? (see image attached) b) Identify 2 possible reasons why this problem is occurring, and for each reason, suggest an experimental modifica on you can do to troubleshoot.
- protein is purified and at a concentration of 600 μg in 1.75 ml of buffer. You do an assay with your purified protein, and the assay requires 25 μg of biochemisfunase. What volume of the purified biochemisfunase would yield 25 μg? Show your calculation.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?You perform a Bradford assay to determine the concentration of isolated α-lactalbumin. You use 50 μL of a two-fold diluted solution of α-lactalbumin in the assay. You generate a standard curve with the following equation for the line: y = 0.163x + 0.082. The absorbance of your sample was 0.674 AU. What is the concentration of α-lactalbumin, in mg/mL, in your sample? Give your answer to three significant figures.
- in isolating ribosomes from a yield sample, describe the ideal type of centrifugation for this separation technique based on the following:*Main type of centrifuge *Subtype centrifuge * Speed range *Operational temperature range*Type of CentrifugationIn a gel filtration chromatography, what type of gel must be used when the protein size is 2500 Da? Explain.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…