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- The colony forming units (CFU)/ml in sauerkraut brine are given for different types of bacteria except for leuconostoc (shaded column). To obtain the Leuconostoc counts for different days, 40 microlitre of sauerkraut brine was 10-fold diluted and the CFU was determined by plating different dilutions on agar plates. For Day 0 56 CFU were counted in 10-1 dilution, Day 1 81 CFU were counted in 10-3 dilution, Day 2 86 CFU were counted in 10-3 dilution Day 5 78 CFU were counted in 10-5 dilution Calculate the CFU/ml of leuconostoc in the brine for the 4 different days. Show your calculationYou measure your E. coli and find them to have an OD600 of 1.20. You have 50 ml of culture, and you know that there are 2.5x109 cells per 1.0 OD600. Based on this information, how many E. coli cells are present in the culture you’re about to centrifuge?Three different strains of Escherichia coli, respectively A, B, and C have been inoculated in three different media, namely I, II, and III, and the growth of each bacterial strain has been followed by spectrophotometry (Figure 1). Strain A Strain B Strain C 4.5 45 Medium I 4 3.5 25 2.5 3 E 2.5 Minimal medium + Glucose 25 2.5 O 15 1 0.5 as 05 100 200 300 400 500 600 100 200 400 600 200 Time (minj Strain A Strain B StrainB 4.5 4s 4s Medium II 3.5 15 25 2 3 Minimal 25 2 medium + Glucose 1 + Arginine 0.5 as as 100 200 300 400 500 600 100 300 400 s00 600 100 300 400 600 Tme Time (min) Strain A me StreinB Streine 4.5 45 45 Medium II 4 4. 3.5 3.5 25 2 3 Minimal medium + Glucose 5 2.5 25 25 8 15 15 + Lactose 1 0.5 05 05 100 200 300 400 500 600 300 200 400 10 200 300 630 Time (min) Tine imin Time min Figure 1. Growth curves of the three E. coli strains A, B and C in three media: I, minimal medium containing glucose; II, minimal medium containing glucose and arginine; and III, minimal medium…
- 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?A serial dilution of overnight E.coli culture was performed by pipetting 1ml of a bacterial culture into a 9 ml LB medium. After this, from 10-4 and 10-5 dilution tubes 100µl were plated onto LB agar plates. Upon overnight incubation at 37°C, 200 colonies were counted in 10-4 and 22 colonies were present on 10-5 plates. How many colony-forming units were present per ml of the original culture? If the formula for CFU/ml =no. Of colonies/dilution factor*volume of culture plateuse these OD numbers to plot growth curve for E. coli K12- and estimate generation time for this culture. Table 1. Absorbance (O.D.600) measured for growing culture of E. coli K12 (time course) Incubation time O.D.600 Incubation time O.D.600 O min 3 hrs 3 hrs 30 min 4 hrs 4 hrs 30 min 5 hrs 5 hrs 30 min 0.11 1.75 30 min 0.20 1.94 1 hr 1 hr 30 min 2 hrs 2 hrs 30 min 0.27 2.24 0.51 2.48 2.31 2.19 0.65 1.30
- Escherichia coli Mycobacterium phlei Bacillus Micrococcus subtilis luteus A B Figure 1: Agar Slant Cultures of Bacteria (Gary E. Kaiser, Ph.D.- Professor of Microbiology) . Observe and describe the colour of the slant cultures (A-D) in fig 1. ( . Define the following terms: pure culture, sterile medium, inoculum, aseptic technique, and colony. What is the name of the cultures that you used . Where they gram negative or positive Define the following terms: psychrophile, mesophile, thermophile, and hyperthermophile.1. Following the modified protocol for the isolation of Escherichia coli bacteriophage of Encabo (2018) presented below, compute for the pfu/ml of the chloroform-treated lysate. 1ml 1ml 1ml 1ml 1ml | | ||| 9ml 9ml 9ml 9ml 9ml Chloroform-treated lysate 101 102 103 104 105 0.1 ml 0.1 ml_0.1 ml Empty sterile tubes ||| + 0.5 ml E. coli Incubate inside ref for 15-20min + 0.1 ml lysate-E.coli mix Molten soft agar overlay O00000 Bottom agar Number of plaque-forming units (pfu) Plate A Dilution Plate B 103 104 105 254 265 132 11 110 23 average number of plaques x dilution factor volume plated pfu mlN. benthamiana will be infiltrated with a solution containing OD600=0.3 of each experimental Agro containing construct and OD600=0.1 of p19. Calculate the volume of cultures (V construct; V p19) needed according to the formulas: V construct = V final × 0.3/OD600; V p19 = V final × 0.1/OD600. One mL of infiltrate is often enough to complete a small experiment. How to plan your final volume accordingly?
- How would you produce a 10^-1 dilution if a 3 mL bacterial sample using the entire 3 mL volume? suppose your professor handed you a test tube with 2.0 mL of an E. coli broth culture in it and told you to make a 10^-1 dilution of the entire culture. Explain how you would do this. Show your calculations.A pure bacterial culture of unknown concentration was diluted to determine the concentration of viable bacteria in the original culture. Serial dilutions were performed as 2. diagrammed below. A volume of 500 µl was transferred into each tube. TSA plates were inoculated with 100 µl from the last three dilution tubes. a. If the dilution between each tube is 102, what is the volume of diluent in each of the 5 dilution tubes? Provide the volume using ml as the units. b. What is the total dilution of tube number 4? Express the total dilution using scientific notation. c. What is the concentration of viable bacteria in the original culture? Express the concentration using scientific notation and CFU/ml as the units. d. If you inoculated a TSA plate with 1.0 ml from dilution tube 4, how many colonies would you expect to form on the plate after incubation? e. If the original culture had a volume of 50ml, what was the total number of viable bacteria in the 50 ml of the original culture? 1 2 3…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 100ul
