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Lab 7 Microbial Genetics & Genetic Engineering BIO250L”

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.

“Pre-Lab Questions”

1. Which DNA nitrogenous bases pair with each other? Which bases are purines, and which are pyrimidines?
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1. How is DNA information used to make proteins? What are the steps of this process?
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2. 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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3. 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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4. 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?

a. Salt (in the DNA extraction solution): Click here to enter text.

b. Detergent (in the DNA extraction solution): Click here to enter text.

c. 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

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Plasmid

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Post-Lab Questions

1. What is the expected size of the plasmid plus the cut foreign DNA?
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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?
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4. How would you clone a gene into a plasmid if there were no common restriction sites between the two DNA sequences?
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