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Prepare a simple flowchart of the experimental procedures for Bacteria Preparation and Transformation
the notes are attached below
Step by step
Solved in 2 steps with 2 images
- What is PCR? Why does Taq polymerase work better than a typical DNA Polymerase isolated from E. coli for PCR? The optimal growth temperature for E Coli is 37 °C.Why is the DNA polymerase used in PCR derived from an extreme thermophile bacteria species rather than a mesophile bacteria species? (be sure to describe the mesophile DNA polymerase as well as the extreme thermophile DNA polymerase.)Currently I am working with qPCR quantification specific for Lactobacillus bacteria. I found in some cases, the sample showing multiple peaks of melting curves while others looks perfect, so do the amplification plot duplication. Any one have an opinion about this issues? Does it caused by sample DNA quality, the omount of the bacteria detected, or primers problem? I am condisering that this condition reflecting the low abundance of the specific bacteria inside its sample.
- At what stage of the culture should bacterial colonies be harvested for plasmid DNA extraction? How about for genomic DNA extraction? What distinguishes the xanthogenate-based methodology from the traditional phenol/chloroform method for isolating DNA from bacteria? What makes potassium ethyl xanthogenate efficient in isolating DNA from a variety of microorganisms?Is the DNA you extracted is pure? What are the possible impurities? What can we do with the DNA once we have purified it? Discuss different techniques and technologies associated with this. Imagine that there is an E. coli outbreak in your area, and you would like to test the kangkong from your local grocery store. How could you modify this protocol to extract DNA from the kangkong (to identify the species) and check for presence or absence of E. coli.? Keep in mind that (i) E. coli is free-living and not an endosymbiont, and (ii) plant cells are encased in both a cell membrane and cell wall.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.
- The following image is of an agarose gel. If DNA samples were loaded to this gel and the electrophoresis experiment was started, explain what would happen and why.TRY TO KEEP IN SHORT AND USE OWN WORD FOR THIS QUESTION You are studying a type of bacteria isolated from the acidic water runoff of a mining operation. You subject two batches of the same bacteria type to different environmental growth conditions. One batch is grown at pH 2, while the other is grown at pH 7. All other environmental parameters are kept identical between the two batches. You then collect their proteins and run a Western blot using an antibody that binds to a proton efflux pump protein (which actively expends energy to pump protons out of a cell). How would you characterize the information obtained in this experiment? What does it tell you, and why is that potentially valuable information?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?
- In bacterial transformation, the purpose of having antibiotic within an agar plate is to: Select one: confirm which plasmids been have successfully ligated with a gene of interest. isolate bacteria which have been successfully transformed with the plasmid. indicate which plasmids were successfully digested by the endonuclease. act as a substrate which will be cleaved and produce a blue product when ligation is unsuccessful. show which plasmids contain the lacZ gene.A graduate student was assaying LD50 (lethal dose 50%) of two temperature-sensitive Francisella tularensis strains in HeLa cells (human cell line). Both strains can infect humans and cause fatal tularemia if untreated, but it is difficult to obtain LD50 values in human subjects. The data below shows LD50 (lethal dose 50%) values of the strains in human cell culture. Can you predict the more virulent strain of the two human pathogens? Francisella tularensis strain A: LD50 @ 20°C= 100; LD50 @ 37°C= 1000 Francisella tularensis strain B: LD50 @ 20°C= 1000 LD 50 @ 37°C= 100 O It is not possible to determine the virulence of the two strains as human pathogens from the provided data Strain A and strain B are equally virulent as human pathogens, as they average out in virulence. O Strain A is more virulent than strain A as a human pathogen. O Strain B is more virulent than strain A as a human pathogen.A graduate student was assaying LD50 (lethal dose 50%) of two temperature-sensitive Francisella tularensis strains in HeLa cells (human cell line). Both strains can infect humans and cause fatal tularemia if untreated, but it is difficult to obtain LD50 values in human subjects. The data below shows LD50 (lethal dose 50%) values of the strains in human cell culture. Can you predict the more virulent strain of the two human pathogens? Francisella tularensis strain A: LD50 @ 20∘C= 100; LD50 @ 37∘C= 1000 Francisella tularensis strain B: LD50 @ 20∘C= 1000 LD50 @ 37∘C= 100 Group of answer choices It is not possible to determine the virulence of the two strains as human pathogens from the provided data Strain A and strain B are equally virulent as human pathogens, as they average out in virulence. Strain A is more virulent than strain A as a human pathogen. Strain B is more virulent than strain A as a human pathogen.