The data below were obtained for the growth of a pure culture of Escherichia coli in nutrient broth at a temperature of 37°C. Determine (1) the specific growth rate and (2) generation time of E. coli and the duration of the (3) lag and (4) log (or exponential) phases. (ln 2 = 0.693) Time (h) 0 1 2 3 4 8 16 32 Bacterial No./mL 104.1 103.9 104.4 105.5 106.5 107.7 108.0 107.6
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The data below were obtained for the growth of a pure culture of Escherichia coli in nutrient broth at a temperature of 37°C.
Determine (1) the specific growth rate and (2) generation time of E. coli and the duration of the (3) lag and (4) log (or exponential) phases. (ln 2 = 0.693)
Time (h) 0 1 2 3 4 8 16 32
Bacterial No./mL 104.1 103.9 104.4 105.5 106.5 107.7 108.0 107.6
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- E. coli has a doubling time of 0.345 hours at 37 0C. Starting with 1 milligram of cells, assuming a lag phase of 30 minutes and a stationary phase of 30 minutes, what number of cells would you expect at the end of 13h, assuming that besides the lag and stationary phases the culture is continuously in exponential phase. Assume that ln (2) = 0.69 and that the mass of a single cell is 10^(-12) g.Two flasks of E. coli are grown in batch culture in the same medium (2% glucose and amino acids; no nitrate) and at the same temperature (378C). Culture #1 is well aerated. Culture #2 is anoxic. After 16 hours the following observations are made: ■ Culture #1 has a high cell density; the cells appear to be in stationary phase, and the glucose level in the medium is reduced to 1.2%. ■ Culture #2 has a low cell density; the cells appear to be in logarithmic phase, although their doubling time is prolonged (over 1 hour). The glucose level is reduced to 0.2%. Why does culture #2 have so little glucose remaining relative to culture #1, even though culture #2 displayed slower growth and has less biomass?Nutrient Agar (NA) is a general purpose medium used for the cultivation of a wide variety of non- fastidious microorganisms (Merck, 2000). Its ingredients are listed in Table 2. You are tasked to prepare 300ml of NA, determine the amount of ingredients and write in column 3 in Table 2. Show your calculations. *A sample calculation was made for you. Peptone = 300 ml x 0.5% = 1.5 g Table 2. Medium Composition of Nutrient Agar (NA) with the recommended proportions (Merck, 2019) Ingredients In % In Grams/300ml Peptone 0.5 *1.5 Meat extract 0.3 Agar 1.5 Distilled Water As needed
- In growing E.coli, why is that (reasons) they do not grow after doubling time under 20 degrees celsius and under 37 degrees celsius, the number of colonies after doubling time has decreased? provide reliable sources (links).in a clean, non-sterile 15 mL centrifuge tube, prepare a 2.0% yeast suspension by adding 0.06 g Saccharomyces cerevisiae to 3 mL yeast growing medium (56 mM glucose, 20 mM HEPES, pH 6.8). What percent of yeast suspension is left after a 1:10 dilution?Nicotiana tabacum cells are cultured to produce a polysaccharide gum. The reactor used is a stirred-tank reactor with an initial volume of 100 L. The maximum specific rate of growth of the culture is 0.18 d-1 and the yield coefficient of substrate in biomass is 0.5 gX/gS. The concentration of the limiting substrate in the medium is 3% (m/v). The reactor is inoculated with 1.5 g/L of cells and operated in batch until the substrate is exhausted, when the medium is fed with a constant flow rate of 8 L/d. The fed batch occurs in a quasi-steady state condition.a) Estimate the time of the batch step and the concentration of cells reached in this phase, considering exponential cell growth.b) The fed batch phase is carried out for a period of 40 days. What is the final concentration of cells in the reactor?c) The bioreactor is available for the process for 275 days a year, with an interval of 24 hours between each cultivation. What is the most advantageous operating mode for the process…
- The nutrient broth is a basic media used for growing a broad variety of microorganisms in the laboratory. The nutrient broth consists mainly of 1.5g * L ^ - 1 extract, 3g * L ^ - 1 yeast extract and 5g * L ^ - 1 sodium chloride dissolved in distilled water. The nutrient agar is prepared by adding the agar at the desired amount into nutrient broth. Answer the following questions. marks) a) How would you prepare a 500 ml nutrient broth? b) How would you prepare 2% (w / v) nutrient agar for 1L? c) How would you prepare a 100 ml nutrient broth supplemented with 100mu * g / m * l ampicillin antibiotic? The stock concentration for ampicillin solution is 100mg / m * l .Bacterial generation times for four different bacterial species were calculated in the media listed in Table 6.3. All media were prepared with pure distilled water and incubated aerobically in the light. Compare and contrast the growth requirements of the four bacteria listed above. Which of the media, if any, are chemically defined?| Generation Time Escherichia coli Pseudomonas Lactobacillus Nitrobacter Medium aeruginos a NaCl, NO3", MgSO4 80 Glucose 100 Glucose, NaCl, PO43- Glucose, NaCl, PO43-, MgSO4 Glucose, NaCl, PO4-, MgSO4, 56 200 43 100 28 40 8 amino acids Glucose, NaCl, PO43-, MgSO4, 25 25 80 19 amino addsYou have just measured the OD of your E. coli (which is in log-phase growth) and it is at 0.15. You are doing a procedure that requires you to centrifuge E.coli once it reaches an OD of 1.20 do you have time to run to the bathroom? In other words, (i) explain the logic, and (ii) show the math to illustrate how you know what time you need to return to lab to centrifuge those bacterial cells.
- 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 100ulSEPARATION OF COMPONENTS BY CENTRIFUGATION Consider the following subcellular components and their corresponding sedimentation rates. Table 3. Sedimentation rates of four subcellular components for isolation. CENTRIFUGAL FORCE (xg) SUBCELLULAR COMPONENT CDCP 14896 AJRA 967 AGC 382 JCKSP 2378 Two centrifuge machines are available in your laboratory. Centrifuge A has a rotor diameter of 14 cm and a maximum speed of 3500 rpm while centrifuge B has a rotor diameter of 18 cm and a maximum speed of 14500 rpm. Construct a separation scheme of the four subcellular components considering the following parameters: The four components are separated into pellet/supernatant I and pellet/supernatant II. Only two centrifugation steps are performed; specify the centrifugal speeds (in rpm) per step. Show relevant calculations. Both centrifuge machines must be used in the procedure. Components of each fraction are indicated.Sydney Brenner isolated Salmonella typhimurium mutants that were implicated in the biosynthesis of tryptophan and would not grow on minimal medium. When these bacterial mutants were tested on minimal medium to which one of four compounds (indole glycerol phosphate, indole, anthranilic acid, and tryptophan) had been added, the growth responses shown in the following table were obtained. Mutant Minimal medium Anthranilic acid Indole glycerol phosphate Indole Tryptophan trp-1 − − − − + trp-2 − − + + + trp-3 − − − + + trp-4 − − + + + trp-6 − − − − + trp-7 − − − − + trp-8 − + − − + trp-9 − − − − + trp-10 − − − − + trp-11 − − − − + Give the order of indole glycerol phosphate, indole, anthranilic acid, and tryptophan in a biochemical pathway leading to the synthesis of tryptophan. Indicate which step in the pathway is affected by each of the mutations.