Imagine you have been given a liquid culture of yeast with a starting concentration of 3.67 x 10' cells/ml and are asked to carry out the sample dilution process shown in the figure below. 100μl 100μl 100μl 100μl 100μl 0.9ml 0.9ml 0.9ml H2O H₂O 6.9ml 0.9ml H₂O H₂O H₂O Original 10-1 102 10-3 104 Culture 105 100μl 100μl 100μl Plate A Plate B Plate C a. How many colonies should have been present on Plate A in this example? - Answers must be whole numbers as partial colonies are not expected. b. Imagine you carried out the same dilution scheme shown in the figure above, but now, you do not know the concentration of the original culture. If you counted 163 colonies on Plate B, what is the concentration of cells/ml in the original culture?
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- A culture of S. cerevisea has an overnight OD of 2.3 (1.0 OD is approx 1.0x107 cells/ml) You will be plating 100µl onto agar and want the final count of colonies on the plate to be around 300 colonies. How much of the 2.3 OD culture must you use to get a 500µl subdilution (with sterile water), so that you have diluted enough to get approx 300 colonies per 100ulTime point (min) Absorbance of culture at 660nm Approximate cell concentration Approximate # cells in 1mL extract 0 0.298 1.49 x 108 cells/mL 1.49 x 108 cells 10 0.316 1.58 x 108 cells/mL 1.58 x 108 cells 20 0.374 1.87 x 108 cells/mL 1.87 x 108 cells 30 0.429 2.145 x 108 cells/mL 2.145 x 108 cells 40 0.512 2.56 x 108 cells/mL 2.56 x 108 cells 50 0.544 2.72 x 108 cells/mL 2.72 x 108 cells 60 0.607 3.035 x 108 cells/mL 3.035 x 108 cells a. Using these data, prepare a growth curve of this strain ofEscherichia coli (E. coli).b. Estimate the doubling time for this strain of E. Coli. Clearly showhow you estimated this value from the empirical data presented.Look at the table below and answer the following questions: Culture 1/10 1/50 1/100 1/200 dilution OD Reading 1 2.0 0.36 0.18 0.11 OD Reading 2 1.9 0.35 0.18 0.08 OD Reading 3 1.8 0.33 0.16 0.09 1. Which 2 dilution levels of the culture are the best to use to compute for its optical density? 2. What is the average optical density of the culture? (use values from 2 dilution levels)
- Describe how you would prepare a dilution series of a 1 x 107 CFU/mL culture to the 10-8 dilution using only 4.5 mL diluents in tubes. What would be the theoretical cell count if 0.1 mL of the 2nd and 4th dilutions were each plated?You prepared a 7x 10^5x dilution from your bacterial culture, plated 0.2 ml of it on a Petridish and counted 67 cfu. What was the cell density of your bacterial culture (in cfu/ml? How many cells did you have in total if the volume of your culture was 50ml? Round to a whole number, do not write in scientific notation. The cell density of my bacterial culture was cfu/ml. The total number of cells wasa. Let’s say for example that a milk sample has 10,000 bacteria per milliliters. If 1 mL of this sample were plated out, these would theoretically be 10,000 colonies in the Petri plate. Discuss and explain the serial dilutions of this example. b. The disk diffusion method was used to evaluate 3 disinfectants. The results were as follows: Solution Zone of inhibition X 0 mm Y 5 mm Z 10 mm How would you interpret? Provide inference.
- You have 0.05 ml of kombucha that contains microbes at a concentration of 8.2 x 10^6 CFU/ml. You then add 4.95 ml fresh tea. What is the dilution? What is the final concentration of cells? Show your calculations.The number of cells in a culture is estimated based on turbidity. If the following standards are used for comparison: Number of cells/mL Optical Density (measure of turbidity) 0.025 0.050 000'000'৮ 000 000'000' కరరక How many cells are present in a culture that has an optical density of 0.075? O A. Less than 1,000,000 O B. Between 2,000,000 and 4,000,000 O C. 16,000,000 O D. Between 4,000,000 and 8,000,000 Reset SelectionIf 0.1 ml of a 1 * 10−6 dilution plate contains 56 colonies, calculate thenumber of cells per ml of the original culture
- A bacterial culture has a concentration of 3.2 x 108 cells /mL. You dilute this culture as follows: 1/50, then 10-3 and finally 1/20. If you then plate 0.2 mL of the final dilution, how many CFU would you expect following incubation?If you have a 15 mg/100 ml stock solution of GA3 and you need a 1 mg GA3 in 25 ml, how much stock solution would you add to 125 ml of medium? how to calculate these kind of question in tissue culture media preperationThe number of bacterial cells in a culture broth is to be determined by a culture technique. Serial dilutions were performed and a 1 mL aliquot from each dilution was mixed with warm molten agar and poured into a Petri dish. The numbers of bacterial colony forming units (CFU) after overnight incubation are shown in the table below. What is the number of colony forming units per mL of the culture broth? Choose only the most appropriate plate for your calculation. Give your answer as the number only (do not add text for the units). You may use scientific notation with the format 1.12e+6 (that is, 1.12 x 106 cfu/mL). (Note: Canvas will then display your answer a whole number.) Plate Dilution Plate 1 10 dilution Plate 2 10 dilution -5 Plate 3 10 dilution Plate 4 10 dilution Plate 5 107 dilution -8 Plate 6 100 dilution *Too many to count Number of colony forming units (CFU) TMTC* TMTC* TMTC* 867 154 18