Microbial Genetics and Genetic Engineering

Student Name: Click here to enter text.                     

Access Code (located on the underside of the lid of your lab kit): Click here to enter text.

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Pre-Lab Questions

  1. Which DNA nitrogenous bases pair with each other? Which bases are purines, and which are pyrimidines?
    Click here to enter text.
  2. How is DNA information used to make proteins? What are the steps of this process?
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  3. Give an example of a scenario in which you would perform PCR vs a scenario in which you would use recombinant DNA technology.
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  4. What occurs during each of the three steps involved in the PCR cycle? How has the use of PCR changed biotechnology?
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  5. How could you take a protein with a known sequence of amino acids and use it to create an artificial gene?
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EXPERIMENT 1: DNA Extraction

Post-Lab Questions

  1. What is the purpose of the following reagents in the experiment?
    1. Salt (in the DNA extraction solution): Click here to enter text.
    2. Detergent (in the DNA extraction solution): Click here to enter text.
    3. Ethanol: Click here to enter text.
  2. What else might be in the ethanol/aqueous interface? How could you eliminate this?
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  3. What is the texture and consistency of the DNA?
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  4. Is the DNA soluble in the aqueous solution or in the alcohol?
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Insert a photo of your DNA. Include your name and access code handwritten in the background of your photo.

 

EXPERIMENT 2: Cloning a DNA Fragment to a Bacterially-Derived Plasmid Vector

Data Tables

Table 1: Fragment Lengths

DNA Type Longest Length (in base pairs)
Foreign Click here to enter text.
Plasmid Click here to enter text.

 

Post-Lab Questions

  1. What is the expected size of the plasmid plus the cut foreign DNA?
    Click here to enter text.
  2. What type of ends do the enzymes BamHI and EcoRI produce? How does this type of end facilitate cloning?
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  3. What enzyme is necessary to permanently link the digested foreign and plasmid DNA together to form the recombinant DNA molecule? How does this enzyme work?
    Click here to enter text.
  4. How would you clone a gene into a plasmid if there were no common restriction sites between the two DNA sequences?
    Click here to enter text.

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