In bacterial transformation ... 1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA? 2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells? 3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes
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In bacterial transformation ...
1, Why do you have to incubate the competent bacterial cells on ice after the addition of plasmid DNA?
2. Why do you have to add SOC (Super Optimal Broth with Catabolite repression) medium to bacterial cells?
3. Why do you have to incubate the bacterial cells in SOC medium at 37oC for 45 minutes?
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- blaA ori lacl Plac rrgB rrgA lacZYA Recombinant Plasmid "B"In E. coli, Base excision repair (BER) handles non-bulky lesions (e.g., deamination, oxidation) and nucleotide excision repair (NER) handles bulky lesions (e.g., Py dimers). Is this statement true or false? O True FalseYou have two DNA samples. Only one was expected to have plasmid DNA that was isolated from it. Both DNA samples were exposed to two different treatment conditions: untreated, or treated with DNase I. How should the DNase I treatment change how DNA appears on the gel?
- When plasmids are isolated from bacterial cells, they may existin a number of forms.a. List the different forms that may be found.b. Which do you think would migrate the fastest and farthest in anelectrophoresis experiment and why?In relation to DNA Isolation experiment 1. Once the tissue has been ground and heated to 60°C, the DNA is released into the solution, but so are many other types of cellular molecules. List some types of molecules besides DNA that you would expect to find in a cell. 2. What is the role of the detergent in this protocol? How does it perform this function? 3. Name two important functions of the proteases? 4. If you wanted to isolate a copy of the gene that codes for a protein found in the skin, could that gene be located in liver cells? Explain your reasoning 5. What do you think will be the first step in purifying DNA from intact isolated cells? 6. What happened to the other macromolecules once the DNA was precipitated out of solution?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?
- How can i explain my answers in depth in microbology for example this question: Assume that there are horizontal gene transfers between two completely different bacterial species. In one case it is a plasmid that is transferred via conjugation, in the other case it is a part of the bacterial chromosome that is transferred via transformation. In which of the two cases is it most likely that the transferred DNA will remain and be able to function in the recipient cells? Explain the biological background to your answer . How do I break down the question so I answer it fully4- Some/ one application(s) for DNA are/ is.... and they/ it start(s) from the mRNA. Therefore, the reverse transcriptase enzyme is required: a) DNA sequencing b) Electrophoresis c) Microarray d) All of the aboveYou have just carried out a transformation using a plasmid (possessing a Amp-resistance gene) that was 0.001 ug/ul in concentration. You added 10 ul of this plasmid to 100 ul of a bacterial cell suspension. Then you added 250 ul of LB following heat shock. After plating 200 ul of this cell suspension with LB onto a LBA plate, you observe 8 colonies on this plate. What is the transformation efficiency of this plasmid (in colonies/ug of plasmid) based on these results? about 1400 O about 80O about 4000 O about 400O O about 8000 f6 米 IO fs fg f1o 19 08. O OO
- When DNA probes are used to detect specific sequence similarities in bacterial DNA, the probe is heated and the two strands of DNA are separated. Why must the probe DNA be heated?Explain Avery-MacLeod-McCarty experiment (the 1940's) with capsule forming bacteria (smooth) and non-capsule forming bacteria (rough). What did this classic experiment demonstrate?Why would a rapidly growing bacterial cell be killed by an antibiotic that blocks DNA replication? Suppose the bacteria form a biofilm on a catheter in a patient. Why might the bacteria have become resistant to the antibiotic?The competency of bacterial cells to take up plasmids from the environment can be enhanced by treating them with calcium chloride. Which of the following statements is true regarding this process? Question 23 options: Chloride ions neutralize charges on the phospholipids and DNA. Chloride ions adhere to the cell membrane and calcium ions to the plasmid DNA, thus increasing the attractive force between them. The calcium ions change the structure of the cell membrane and, as a result, the pores are enlarged. Calcium ions neutralize charges on the phospholipids of the bacterial cell membrane and on the DNA of the plasmid. Chloride ions enter the cell through protein pores in the membrane, carrying plasmid DNA with them.