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- You are setting up a restriction digest of pUC19. The DNA concentration is 76.4ng/ul. You want to cut with Ndel and Kasl. How would you set up this reaction?HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952 publication by Hershey and Chase. Bacteriophage were labeled with radioactive tracers and allowed 10 infect bacteria. The virusbacteria mixtures were then whirled in a blender to dislodge any viral components attached to the exterior of the bacteria. Afterward, radioactivity from the tracers was measured. FIGURE 8.5 Detail of Alfred Hershey and Martha Chases 1952 publication describing their experiments with bacteriophage. Infected bacteria refers to the percentage of bacteria that survived the blender. How did the researchers know that the radioisotopes in the fluid came from outside of the bacterial cells and not from bacteria that had been broken apart by whirling in the blender?HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952 publication by Hershey and Chase. Bacteriophage were labeled with radioactive tracers and allowed 10 infect bacteria. The virusbacteria mixtures were then whirled in a blender to dislodge any viral components attached to the exterior of the bacteria. Afterward, radioactivity from the tracers was measured. FIGURE 8.5 Detail of Alfred Hershey and Martha Chases 1952 publication describing their experiments with bacteriophage. Infected bacteria refers to the percentage of bacteria that survived the blender. After 4 minutes in the blender, what percentage of each isotope was extracellular?
- HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952 publication by Hershey and Chase. Bacteriophage were labeled with radioactive tracers and allowed 10 infect bacteria. The virusbacteria mixtures were then whirled in a blender to dislodge any viral components attached to the exterior of the bacteria. Afterward, radioactivity from the tracers was measured. FIGURE 8.5 Detail of Alfred Hershey and Martha Chases 1952 publication describing their experiments with bacteriophage. Infected bacteria refers to the percentage of bacteria that survived the blender. The extracellular concentration of which isotope increased the most with blending?HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952 publication by Hershey and Chase. Bacteriophage were labeled with radioactive tracers and allowed 10 infect bacteria. The virusbacteria mixtures were then whirled in a blender to dislodge any viral components attached to the exterior of the bacteria. Afterward, radioactivity from the tracers was measured. FIGURE 8.5 Detail of Alfred Hershey and Martha Chases 1952 publication describing their experiments with bacteriophage. Infected bacteria refers to the percentage of bacteria that survived the blender. Before blending what percentage of each isotope. 35S and 32P, was extracellular (outside the bacteria)?HersheyChase Experiments The graph shown in FIGURE 8.5 is reproduced from an original 1952 publication by Hershey and Chase. Bacteriophage were labeled with radioactive tracers and allowed 10 infect bacteria. The virusbacteria mixtures were then whirled in a blender to dislodge any viral components attached to the exterior of the bacteria. Afterward, radioactivity from the tracers was measured. FIGURE 8.5 Detail of Alfred Hershey and Martha Chases 1952 publication describing their experiments with bacteriophage. Infected bacteria refers to the percentage of bacteria that survived the blender. Do these results imply that viruses inject DNA or protein into bacteria? Why or why not?
- 1. If you do an absorbance reading after plasmid purification and get a A260/A280 of less than 1.8, how could you further purify the sample to get rid of the protein contamination? Is it always necessary to have completely pure DNA? What are some cases where it would or would not be?5 μL of plasmid DNA (pUC 19) was added to 50ul of CaCl2 competent E. coli that was then added 250 μL of SOC medium, 100 μL of this solution was plated onto a TSA + ampicillin plate. Can you please show me how I calculate the total number of successful transformants per mL of competent cells plated. The total number of colnies counted on plate was 70. Thank youYou overexpressed (asked bacteria to make a lot of) an 80 kDa protein carrying a 6X-His tag and purified it on a nickel column. You collect frac ons from your purifica on and analyze them using SDSPAGE. The results are shown in image attached. a) Interpret the results from the gel. What is wrong? (see image attached) b) Identify 2 possible reasons why this problem is occurring, and for each reason, suggest an experimental modifica on you can do to troubleshoot.
- 1a)What is the purpose of adding 10mM of Tris-HCL (pH 7.5) during the extraction process? Oa. It breaks down the cell membrane to release the DNA. Ob. It ensures that the DNA that is released from the cell is turned into single stranded format. Oc. It neutralizes the the lysis reaction. Od. It denatures the proteins that were released from the cell.1b) What is the purpose of the Phosphate Buffered Saline (PBS) in the DNA extraction protocol? O a. The swab containing cheek cells is swirled around in this buffer to collect the cells. O b. It is used to breakdown the cell membrane to release the DNA. O c. It is used to denature proteins that were released from the cell. Od. It is used as the main buffer in the PCR reaction.Explain 2 possible consequences of accidentally puncturing one of the wells in the agarose gel in an electrophoresis set-up when loading a DNA sample.1. Labels are used in biosensors to amplify a biorecognition event. Which is frequently used in lateral flow assays? C-13 or P-32 nanoparticles fluorophores labeled enzymes 2. Why is it a good idea to use filters? Because of the mains interference All of these Because of the parasitic Because the signal is mixed with other unwanted signals