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You seed another culture of different end cell line, Walker12 cells, at 8 am on Friday with 10^4 cells. By Thursday at noon (12:00 pm) they reach confluence. Assume the sae confluency as stated above for a T150 flask (culture area of 150cm^2 adn when confluent contains 2 x 10^7 cells). What is the generatoin time (doubling time)?
A. 120 hours
B. 80 hours
C. 38 hours
D. 8 hours
E. 11.4 hours
F. other ________
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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?An Hfr strain that is leuA+ and thiL+ was mixed with a strain thatis leuA− and thiL−. In the data points shown in the following graph,the conjugation was interrupted at different time points, and thepercentage of recombinants for each gene was determined bystreaking on a medium that lacked either leucine or thiamine.What is the map distance (in minutes) between these two genes?Since higher concentration colcemid will result in shorter chromosome, you want to change your protocol and reduce final concentration of colcemid in your 10 ml blood culture from 0.1ug/ml to 0.05ug/ml. how many ul colcemid stock solution with concentration10ug/ml needed to be added in 10ml blood culture?
- A high cell density culture of recombinant E. coli was carried out according to the following strategy:-Step 1: single batch with exponential growth until 98% conversion of the substrate, starting from V0= 4.0 L, S0=50 g/L/ X0= 1.0 g/LStep 2: batch fed with exponential flow (SF-800 g/L, μ= 0.1 h-1) until reaching X= 50.9 g/L;Step 3: batch fed with constant flow (F= 0.1 L/h) for 4 hours (induction phase with IPTG)Note: consider that the quasi-steady state is reached in both fed-batch stages.Extra data: YX/S = 0.4 gx/gs; μmax= 0.25 h-1; Ks== 1.0 g/L a) What was the cell concentration reached at the end of step 1?b) For step 3, considering that the substrate concentration in the feed was 1/4 of that used in step 2, what was the concentration of cells reached at the end of step 3?C) In terms of cell productivity, which of the three phases of cultivation was the most productive?a. Which wells do contain more cell number when compared to other wells in cell viability with resazurin assay?b. Calculate cell viability percentage. 570nM Sample 1 Sample 2 Sample 3 Sample 4 control 0,154 0,157 0,155 0,154 1/1000 0,15 0,149 0,151 0,148 1/100 0,113 0,115 0,116 0,114 1/75 0,052 0,05 0,052 0,051 1/50 0,03 0,031 0,029 0,032 1/25 0,019 0,018 0,021 0,019a sample was found to contain 10.0ug/L of nitrate. a spike of 5 ug/L was added to a replicate unknown, and the spiked sample was found to contain 14.6 ug/L. what is the percent recovery?
- Production of a recombinant protein by E. coli is proportional to cell growth. Ammonia is used as a source of N and glucose as a source of C, under aerobic conditions. The recombinant protein has a general formula CH1.55N0.31O0.25 and the cell CH1.77N0.4900.24. The yield of biomass from glucose is 0.48 gcel/gglic, and the yield of recombinant protein from glucose is about 20% of that for biomass.a) How much ammonia is needed to produce 50 g of cells producing the recombinant protein?b) What is the oxygen demand in this process?c) For the cultivation of a wild E. coli not producing the recombinant protein, how different would the ammonia and oxygen demand be if the biomass yield remained at 0.48 gcel/gglyc ?An Hfr strain that is leuA+ and thiL+ was mixed with a strain thatis leuA− and thiL−. In the data points shown in the following graph,the conjugation was interrupted at different time points, and thepercentage of recombinants for each gene was determined bystreaking on a medium that lacked either leucine or thiamine.What information do you know based on the question and your understanding of the topic?In Figure 5-5,a. Why do A− and B− cells, by themselves, not formcolonies on the plating medium?b. What genetic event do the purple colonies in themiddle plate represent?
- In the Avery, McLeod, McCarty Experiment where supernatant from heat killed, virulent S Strain pneumonia solutions were added to non-virulent R Strain pneumonia cell cultures and allowed to grow in liquid media (i.e., broth). In tubes where Protease was added to the supernatant prior to cell culture, what was the observed effect when plating and growing the S. pneumonia cells to solid media?In conducting feeding trials to evaluate the effect of feed additives: a. Why is it important to include a negative control in the treatments? b. Why do u need to ensure that the number of replicates or sub-samples are sufficient?You co-culture the following bacterial strains: an Hfr prototroph and an F- auxotroph for the genes mal, met, mtl, and xyl. You interrupt conjugation at various time points and place the mixtures on media plates lacking each of the nutrients. Based on the results shown on the right, what is the order of these four genes along the bacterial chromosome? Explain your reasoning.