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DNA Extraction and Agarose Gel Electrophoresis Questions

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Complete the following tasks.
You discovered that a species of bacteria can break down Styrofoam™ (polystyrene) products due to an
enzyme it produces, polystyrenase. You wish to study the gene that codes for this enzyme.
Task 1: DNA Extraction
To begin work on the bacterium, you begin by extracting its genomic DNA (gDNA). What is the purpose of
the following procedures? Answer briefly but completely.
a. Using sodium dodecyl sulfate, a detergent
Answer: disrupts the cell membrane to release cell contents (and denatures proteins)
b. Adding RNase A and Proteinase K during extraction
Answer: remove/degrade RNA and protein contaminants
c. Adding ethanol before recovering the DNA extract
Answer: precipitate the DNA (by lowering dielectric constant)
Task 2: Polymerase Chain Reaction
After purifying the gDNA extract, you want to isolate and amplify the polystyrenase gene. You perform PCR
using the appropriate gene-targeted primers. What is the purpose of the following PCR components? Answer
briefly but completely.
a. DNA polymerase isolated from Thermus aquaticus
Answer: replicate template DNA even at high temperature (thermostable)
b. Deoxynucleotide triphosphates (dNTPs)
Answer: source of nucleotides incorporated into the new/nascent DNA strands
c. Forward and reverse primers
Answer: determine which segment of DNA (the targeted gene) will be amplified specifically
Task 3: Agarose Gel Electrophoresis
Your amplicon from PCR was subjected to AGE for analysis. You are shown the image of the gel loaded
with the following lanes: (A) negative control, (B) size ladder (1, 2, 3, 4, 6, and 10 kb), (C) gDNA extract, (D)
PCR amplicon. However, due to mishaps while loading the gel with the samples, you are not sure which lane
is which. You are shown a diagram of the obtained gel below.

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Complete the following tasks. You discovered that a species of bacteria can break down Styrofoam™ (polystyrene) products due to an enzyme it produces, polystyrenase. You wish to study the gene that codes for this enzyme. Task 1: DNA Extraction To begin work on the bacterium, you begin by extracting its genomic DNA (gDNA). What is the purpose of the following procedures? Answer briefly but completely. a. Using sodium dodecyl sulfate, a detergent Answer: disrupts the cell membrane to release cell contents (and denatures proteins) b. Adding RNase A and Proteinase K during extraction Answer: re ...
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Anonymous
Very useful material for studying!

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