18. The total number of cells in a culture is counted using the trypan blue exclusion assay and is found to be 2.7 x 10^6 cells/ml. The culture is diluted 1:27 and then 100μl seeded per well into a 96 well plate. What is the final cell density per well? Od) 1 x 10^4 Ob) 2.7 x 10^4 Oc) 2.7 x 10^5 Oa) 1x10^5
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- A.Why do you plate the cells from the viable count on LB agar without ampicillin? B.If you observe 100 colonies on your 1/100 plate, how many colonies do you expect if everything works perfectly on your 1/1000 plate?Diagram illustrating Miles & Misra technique for determining viable counts: 10ul 10-5 106 104 10-7 3. Set up controls of the donor and recipient cultures as follows: • Spread plate 0.1 ml of the donor culture over the surface of each of the 3 different selective media (i.e. nutrient agar + Ap; nutrient agar + Sm; nutrient agar + Rif) • Similarly spread plate 0.1 ml of the recipient culture over the surface of each of the 3 selective media • What is the purpose of this step?At the station there is a 2 mL microcentrifuge tube containing sheep blood and 3 tubes containing the following solutions: 0 mM, 300 mM, 600 mM of sucrose. The tubes are randomly labelled A, B, C. 2 mL of each solution were transferred to a different microcentrifuge tube, 20 ul of blood were then added to each tube, and they were mixed well by inverting the tube for multiple times. They were observed and it was determined whether the solution in each tube was hypotonic (Yes or No) relative to the blood cells. In addition, the solutions were diluted with water to help you determine which solution is which sucrose concentration. What sucrose concentration corresponds with each tube? Tube Hypotonic (Yes/No) Hypotonic after 1:1 dilution with water (Yes/No) Most likely Sucrose concentration (mM) A No Yes B Yes Yes C No No
- There are two cultures of yeast cells in the pictures, one has been incubated for 6 hours and one has been incubated for 24 hours. After a 10x dilution by taking 100µl of each culture and adding it to 900 µl water in a microcentrifuge tube and 100µl sample from the tube was taken to view in the counting chamber. a) Count the total number of yeast cells for each culture respectively b) Calculate the concentration and density of yeast cells for each culture respectivelyDiscuss the role of the addition of ammonium chloride potassium lysius buffer in splenocyte culture preparatrion.A phagehunter performs a full plate titer using standard techniques (100 ul of each dilution added to 250 ul of Gordonia terrae cells) of a lysate obtained from an optimum webbed plate experiment. The phagehunter counts 72 plaques on the 10-7 dilution plate. Which of the following scenarios is the best choice for the phagehunter to do next? a.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should adopt a phage from direct isolation. b.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should go back to the pick-a-plaque experiment. c.) The phagehunter has achieved a high enough titer lysate to move forward with characterization experiments. d.) The phagehunter has not achieved a high enough titer lysate to move forward with characterization experiments, so they should try to make more optimum webbed plates.
- What is the dilution factor in the task given if the total viable cells is 140 [28×5(squares)] and the total nonviable cells is 50 [10x5(squares)]?Why is the plate count method a determination of the number of viable cells?a. Explain whether or not any of the methods in fi gure 2.9 could beused to determine the total number of cells present in a patient’s specimen.b. After performing the streak plate method on a bacterial specimen, theculture was incubated for 48 hours at 37°C. Upon viewing the plate, therewas heavy growth (with no isolated colonies) in the fi rst quadrant, but nogrowth was apparent in the remaining quadrants. Please discuss errors in the procedure that could have produced this result.
- In lab we learned a technique that helped us to visulize individual colonies of bacter 1. Describe this technique. 2. What do you expect the resutls to look like? Be specific. 3. How can this technique help you to determine if your culture is contaminated? For the toolbar, press ALT+F10 (PC) or ALT+FN+F10 (Mac). BIUS Paragraph I +] F H Ix X ABC † ( O K₂ KN Q V Arial sè "Ω Θ A 4 10pt EE 88 A Click Save and Submit to save and submit. Click Save All Answers to save all answers. 描く前 X² X₂ 3 由用目1. Draw and describe the possible result that may be observed on an agar plate that utilized a 4- streak quadrant method for isolation WITHOUT the conduct of proper aseptic techniques Source of inoculum: Tap water Characteristic of colony: Yellow, circular colony with entire margin and convex elevation Answer: 2. Using your fingers, you are asked to aseptically touch the surface of a sterile agar plate. Illustrate the possible result from this step if your fingers are (a) unwashed – touched various things prior to placing on agar surface, and (b) washed with soap or disinfected with 70% alcohol. Describe the relative abundance of microbial growth observed on the plates. Unwashed Washed Answer. 3. List and draw the possible characteristics of an isolated bacterial colony that can be observed based on type of (a) margin, (b) elevation, (C) texture, and (d) optical property.1. Draw and describe the possible result that may be observed on an agar plate that utilized a 4-streak quadrant method for isolation WITHOUT the conduct of proper aseptic techniques Source of inoculum: Tap water Characteristic of colony: Yellow, circular colony with entire margin and convex elevation Answer: 2. Using your fingers, you are asked to aseptically touch the surface of a sterile agar plate. Illustrate the possible result from this step if your fingers are (a) unwashed – touched various things prior to placing on agar surface, and (b) washed with soap or disinfected with 70% alcohol. Describe the relative abundance of microbial growth observed on the plates. Unwashed Washed