you plated E. coli that had been transformed with your ligation reactions. There will also be a positive control that has E coli transformed with PHSG298 and a negative control that had E. coli transformed with water. Each was plated on both LB plates and LB + kanamycin plates. Match the growth pattern you
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- 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?We transformed E coli cells with a plasmid modified to contain a 'virulence factor' which would allow growth on media containing the antibiotic kanamycin (Kan). The plasmid confers constitutive resistance to ampicillin (Amp) Assume you were given competent cells of known transformation efficiency (TE). Assume TE= 1×10[6] (note 10[6] means 10 to the power of 6). You want to have about 1000 colonies on the P-200 plate. How many nanograms of plasmid should you use in the transtormation reaction? О 1.5.00 O 2.0.05 О 3.50.00 О 4.200.00 О 5.20.00 О 6.0.50We transformed E coli cells with a plasmid modified to contain a ‘virulence factor’ which would allow growth on media containing the antibiotic kanamycin (Kan). The plasmid confers constitutive resistance to ampicillin (Amp). Assume you were given competent cells of known transformation efficiency (TE). Assume TE= 1x10[7] (note 10[7] means 10 to the power of 7). You want to have about 1000 colonies on the P-200 plate. How many nanograms of plasmid should you use in the transformation reaction? Select only one answer. 1. 0.05 2. 20.00 3. 0.50 4. 50.00 5. 200.00 6. 5.00
- You generate several temperature sensitive mutant strains of bacteria. To study what genes might be affected, the mutant strains are cultured along with the normal parental strain at the permissive temperature (37°C) and then shifted at 10 minute intervals to the non permissive temperature (42°C). The only nucleotides in the growth environment are labeled with 32P. The amount of DNA at each step is measured by determining the amount of labeled nucleotides that have been incorporated into the DNA. The numbers refer to the relative amount of labeled nucleotides that have been incorporated. The following results are obtained.(picture) Which strains/strains, if any, has a mutation in a gene required for DNA replication? Explain your experimental predictions and the degree to which the data support/fail to support them.For bacteria that are F+, Hfr, F', and F- answer the following. a. Describe the state of the F factor. b. Which of these cells are donors? Which is the recipient? c. Which of these donors can convert exconjugants to a donor state? d. Which of these donors can transfer a donor gene to exconjugants? e. Describe the results of conjugation (i.e., changes in the recipient and the exconjugant) that allow detection of the state of the F factor in a donor strain. f. Describe a "partial diploid" and how it originates.you have transformed E.coli cells with a plasmid containing the gene for the enzyme beta-lactamase you have used 3µl of a stock solution of 2.5ng/µl DNA. You transformed 150/µl of competent cells, using the calcium chloride method. you have added 600µl of luria broth to the cells and then plated out 200µl on triplicate plates. upon counting the colonies, you got the following results: plate 1: 120 cfu plate 2: 35 cfu plate 3: 95 cfu please answer the following question: 1. what new characteristic will the cells have ater transformation? 2. how are you going to test for the new characteristic? 3. calculate the transformation efficiency of the experiment?
- We isolate DNA from a bacteria that is wild type for the genes arg leu and his shear it in a blender mix it into a culture of bacteria that is mutant for the genes arg leu and his . We then plate on various media. These are the results: Media with arginine 20 colonies /ml Media with histidine 300 colonies /ml Media with leucine 500 colonies /ml Media with all three 10000 colonies/ml 21. What is this experiment called? A. transpiration B. conjugation C. transformation D. transductionYou found five T4 rII− mutants that will not grow onE. coli K(λ). You mixed together all possible combinations of two mutants (as indicated in the followingchart), added the mixtures to E. coli K(λ), and scoredfor the ability of the mixtures to grow and makeplaques (indicated as a + in the chart).1 2 3 4 51 − + + − +2 − − + −3 − + −4 − +5 −a. How many genes were identified by this analysis?b. Which mutants belong to the same complementation groups?Select all that apply from the following to each transformation process : The following are White non-glowing colonies White glowing colonies Blue non-glowing colonies Blue glowing colonies None of the above The transformation are: a. LB + kanamycin plates that have been spread with the E. coli cells transformed with your ligations, what phenotype of the colonies do you expect to obtain . b. LB + kanamycin plates that have been spread with the E. coli cells transformed with water, what phenotype of the colonies do you expect to obtain. c. LB plates that have been spread with the E. coli cells transformed with your pHSG298, what phenotype of the colonies do you expect to obtain. d. LB + kanamycin plates that have been spread with the E. coli cells transformed with PHSG298, what phenotype of the colonies do you expect to obtain. e. LB plates that have been spread with the E. coli cells transformed with your ligations, what phenotype of the colonies do you expect to obtain. f. LB plates…
- 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?Results from a Kirby Bauer antibiotic assay on a Gram-negative bacterial culture are described as follows: A) the bacterium is resistant to penicillin, an antibiotic that targets synthesis of the peptidoglycan cell wall and B) the bacterium is resistant to tetracycline, an antibiotic that targets the small subunit of the ribosome, inhibiting protein synthesis. Which of the results represents intrinsic resistant and which represents acquired resistants?Describe how will you obtain a 0.2 U of Pst1 restriction enzyme from a stock solution of 5 U/uL of Psti restriction enzyme, using a P10 micropipette, with 0.5 uL limit of detection.